U.S. patent number 7,414,071 [Application Number 10/386,307] was granted by the patent office on 2008-08-19 for methods of treatment with selective ep4 receptor agonists.
This patent grant is currently assigned to Pfizer Inc.. Invention is credited to Kimberly O. Cameron, Delvin R. Knight, Jr., Bruce A. Lefker.
United States Patent |
7,414,071 |
Cameron , et al. |
August 19, 2008 |
Methods of treatment with selective EP4 receptor agonists
Abstract
The present invention provides a method of treating
hypertension, liver failure, loss of patency of ductus arteriosus,
glaucoma or ocular hypertension in a patient, comprising
administering to the patient a therapeutically effective amount of
a selective EP.sub.4 receptor agonist of Formula I ##STR00001## or
a prodrug thereof, a pharmaceutically acceptable salt of the
selective EP.sub.4 receptor agonist or prodrug or a stereoisomer or
diastereomeric mixture of the EP.sub.4 receptor agonist, prodrug or
salt, wherein the variables X, Z, Q, , and R.sup.2 are as defined
in the specification.
Inventors: |
Cameron; Kimberly O. (East
Lyme, CT), Lefker; Bruce A. (Gales Ferry, CT), Knight,
Jr.; Delvin R. (Ledyard, CT) |
Assignee: |
Pfizer Inc. (New York,
NY)
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Family
ID: |
28042041 |
Appl.
No.: |
10/386,307 |
Filed: |
March 11, 2003 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20030207925 A1 |
Nov 6, 2003 |
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Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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60365711 |
Mar 18, 2002 |
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Current U.S.
Class: |
514/422; 514/438;
514/424; 514/382; 514/381 |
Current CPC
Class: |
C07D
409/06 (20130101); C07D 207/26 (20130101); A61P
9/12 (20180101); A61P 27/06 (20180101); A61K
31/4025 (20130101); C07D 417/06 (20130101); A61P
1/16 (20180101); C07D 403/06 (20130101); A61K
31/4015 (20130101); A61P 43/00 (20180101); A61P
9/10 (20180101); A61K 31/427 (20130101) |
Current International
Class: |
A61K
31/40 (20060101); A61K 31/38 (20060101); A61K
31/41 (20060101) |
Field of
Search: |
;514/438,444-449,422,424,381,382 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
Primary Examiner: Wang; Shengjun
Attorney, Agent or Firm: Zielinski; Bryan C. Hsu; Wendy
L.
Parent Case Text
CROSS-REFERENCE TO RELATED APPLICATION
This application claims priority from U.S. provisional application
No. 60/365,711 filed on Mar. 18, 2002.
Claims
The invention claimed is:
1. A method of treating glaucoma or ocular hypertension in a
patient in need thereof, comprising administering to the patient a
therapeutically effective amount of a selective EP.sub.4 receptor
agonist of Formula I ##STR00024## or a prodrug thereof, a
pharmaceutically acceptable salt of the selective EP.sub.4 receptor
agonist or prodrug or a stereoisomer or diastereomeric mixture of
the EP.sub.4 receptor agonist, prodrug or salt, wherein:
##STR00025## X is --CH.sub.2-- or O; Z is thienyl Q is carboxyl,
(C.sub.1-C.sub.4)alkoxylcarbonyl or tetrazolyl; R.sup.2 is --Ar or
--Ar.sup.1--V--Ar.sup.2; V is a bond, --O--, --OCH.sub.2-- or
--CH.sub.2O--; Ar is a partially saturated, fully saturated or
fully unsaturated five to eight membered ring optionally having one
to four heteroatoms selected independently from oxygen, sulfur and
nitrogen, or a bicyclic ring consisting of two fused independently
partially saturated, fully saturated or fully unsaturated five or
six membered rings, taken independently, optionally having one to
four heteroatoms selected independently from nitrogen, sulfur and
oxygen, said partially or fully saturated ring or bicyclic ring
optionally having one or two oxo groups substituted on carbon or
one or two oxo groups substituted on sulfur; and Ar.sup.1 and
Ar.sup.2 are each independently a partially saturated, fully
saturated or fully unsaturated five to eight membered ring
optionally having one to four heteroatoms selected independently
from oxygen, sulfur and nitrogen, said partially or fully saturated
ring optionally having one or two oxo groups substituted on carbon
or one or two oxo groups substituted on sulfur; said Ar moiety is
optionally substituted on carbon or nitrogen, on one ring if the
moiety is monocyclic, or on one or both rings if the moiety is
bicyclic, with up to three substituents per ring each independently
selected from hydroxy, halo, carboxy, (C.sub.1-C.sub.7)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.7)alkyl, (C.sub.2-C.sub.7)alkenyl,
(C.sub.3-C.sub.7)cycloalkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)- alkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)alkanoyl, formyl,
(C.sub.1-C.sub.8)alkanoyl,
(C.sub.1-C.sub.6)alkanoyl(C.sub.1-C.sub.6)alky- l,
(C.sub.1-C.sub.4)alkanoylamino,
(C.sub.1-C.sub.4)alkoxycarbonylamino, hydroxysulfonyl,
aminocarbonylamino or mono-N--, di-N,N--, di-N,N'-- or
tri-N,N,N'--(C.sub.1-C.sub.4)alkyl substituted aminocarbonylamino,
sulfonamido, (C.sub.1-C.sub.4)alkylsulfonamido, amino, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylamino, carbamoyl, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylcarbamoyl, cyano, thiol,
(C.sub.1-C.sub.6)alkylthio, (C.sub.1-C.sub.6)alkylsulfinyl,
(C.sub.1-C.sub.4)alkylsulfonyl and mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylaminosulfinyl, wherein said alkyl and
alkoxy substituents in the definition of Ar are optionally
substituted on carbon with up to three fluoro atoms; and said
Ar.sup.1 and Ar.sup.2 moieties are independently optionally
substituted on carbon or nitrogen with up to three substituents
each independently selected from hydroxy, halo, carboxy,
(C.sub.1-C.sub.7)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.7)alkyl, (C.sub.2-C.sub.7)alkenyl,
(C.sub.3-C.sub.7)cycloalkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)alkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)alkanoyl, formyl,
(C.sub.1-C.sub.8)alkanoyl,
(C.sub.1-C.sub.6)alkanoyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.4)alkanoylamino,
(C.sub.1-C.sub.4)alkoxycarbonylamino, hydroxysulfonyl,
aminocarbonylamino or mono-N--, di-N,N--, di-N,N-- or
tri-N,N,N--(C.sub.1-C.sub.4)alkyl substituted aminocarbonylamino,
sulfonamido, (C.sub.1-C.sub.4)alkylsulfonamido, amino, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylamino, carbamoyl, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylcarbamoyl, cyano, thiol,
(C.sub.1-C.sub.6)alkylthio, (C.sub.1-C.sub.6)alkylsulfinyl,
(C.sub.1-C.sub.4)alkylsulfonyl and mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylaminosulfinyl, wherein said alkyl and
alkoxy substituents in the definition of Ar.sup.1 and Ar.sup.2 are
optionally substituted on carbon with up to three fluoro atoms.
2. A method of claim 1 wherein the selective EP.sub.4 receptor
agonist is a compound of Formula Ia ##STR00026## R is Ar wherein
said Ar moiety is optionally substituted on carbon or nitrogen, on
one ring if the moiety is monocyclic, or on one or both rings if
the moiety is bicyclic, with up to three substituents per ring each
independently selected from hydroxy, halo, carboxy,
(C.sub.1-C.sub.7)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.7)alkyl, (C.sub.2-C.sub.7)alkenyl,
(C.sub.3-C.sub.7)cycloalkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)alkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)alkanoyl, formyl,
(C.sub.1-C.sub.8)alkanoyl,
(C.sub.1-C.sub.6)alkanoyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.4)alkanoylamino,
(C.sub.1-C.sub.4)alkoxycarbonylamino, hydroxysulfonyl,
aminocarbonylamino or mono-N--, di-N,N--, di-N,N'-- or
tri-N,N,N--(C.sub.1-C.sub.4)alkyl substituted aminocarbonylamino,
sulfonamido, (C.sub.1-C.sub.4)alkylsulfonamido, amino, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylamino, carbamoyl, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylcarbamoyl, cyano, thiol,
(C.sub.1-C.sub.6)alkylthio, (C.sub.1-C.sub.6)alkylsulfinyl,
(C.sub.1-C.sub.4)alkylsulfonyl and mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylaminosulfinyl, wherein said alkyl and
alkoxy substituents in the definition of Ar.sup.1 and Ar.sup.2 are
optionally substituted on carbon with up to three fluoro.
3. A method of claim 2 wherein the selective EP.sub.4 receptor
agonist is a compound of the Formula Ia, wherein Ar is cyclohexyl,
1,3-benzodioxolyl, thienyl, naphthyl or phenyl optionally
substituted with one or two (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl, chloro, fluoro,
trifluoromethyl or cyano, wherein said alkyl and alkoxy
substituents in the definition of Ar are optionally substituted
with up to three fluoro.
4. A method of claim 3 wherein the selective EP.sub.4 receptor
agonist is a compound of the Formula Ia, wherein ##STR00027## is a
single bond; Q is carboxy or (C.sub.1-C.sub.4)alkoxylcarbonyl; and
Z is ##STR00028##
5. A method of claim 4 wherein the selective EP.sub.4 receptor
agonist is a compound of the Formula Ia, a prodrug thereof, a
pharmaceutically acceptable salt of said compound or said prodrug
or a stereoisomer or diastereomeric mixture of said compound,
prodrug or salt, wherein Q is carboxy and Ar is phenyl optionally
substituted with one (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl, chloro, fluoro,
trifluoromethyl or cyano, wherein said alkyl and alkoxy
substituents in the definition of Ar are optionally substituted
with up to three fluoro.
6. A method of claim 5 wherein the selective EP.sub.4 receptor
agonist is a compound of the Formula Ia, a prodrug thereof, a
pharmaceutically acceptable salt of said compound or said prodrug
or a stereoisomer or diastereomeric mixture of said compound,
prodrug or salt, wherein Ar is 3-trifluoromethylphenyl.
7. A method of claim 5 wherein the selective EP.sub.4 receptor
agonist is a compound of the Formula Ia, a prodrug thereof, a
pharmaceutically acceptable salt of said compound or said prodrug
or a stereoisomer or diastereomeric mixture of said compound,
prodrug or salt, wherein Ar is 3-chlorophenyl.
8. A method of claim 5 wherein the selective EP.sub.4 receptor
agonist is a compound of the Formula Ia, a prodrug thereof, a
pharmaceutically acceptable salt of said compound or said prodrug
or a stereoisomer or diastereomeric mixture of said compound,
prodrug or salt, wherein Ar is 3-trifluoromethoxyphenyl.
9. A method of claim 5 wherein the selective EP.sub.4 receptor
agonist is a compound selected from the group consisting of
5-(3-(2S-(3R-hydroxy-4-(3-trifluoromethylphenyl)butyl)-5-oxo-pyrrolidin-1-
-yl)propyl)thiophene-2-carboxylic acid;
5-(3-(2S-(3R-hydroxy-4-(3-trifluoro-methoxyphenyl)butyl)-5-oxo-pyrrolidin-
-1-yl)propyl)thiophene-2-carboxylic acid; and
5-(3-(2S-(4-(3-chloro-phenyl)-3R-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl)pro-
pyl)thiophene-2-carboxylic acid.
Description
FIELD OF THE INVENTION
The present invention relates to methods for the treatment of
disorders responsive to modulation of the prostaglandin E.sub.2
receptor, in a patient in need thereof, by administration of a
receptor selective prostaglandin E.sub.2 agonist. More
specifically, the present invention provides methods for the
treatment of hypertension, liver failure, loss of patency of the
ductus arteriosus, glaucoma or ocular hypertension in a patient in
need thereof by administration of a selective prostaglandin E.sub.2
type 4 receptor agonist.
BACKGROUND OF THE INVENTION
The naturally occurring prostaglandins are comprised of several
biological entities including prostaglandin E (PGE). Prostaglandin
E.sub.2 (abbreviated as PGE.sub.2 herein) is known to be a
cyclooxygenase induced oxidative metabolite in the arachidonic acid
cascade, and it has been well documented that prostaglandins,
including PGE.sub.2, have effects on many of the organs and systems
of the body. For example, it is known that PGE.sub.2 has
cyto-protective activity, uterine contractile activity, a
pain-inducing effect, a promoting effect on digestive peristalsis,
an awakening effect, a sleep-inducing effect, a suppressive effect
on gastric acid secretion, hypotensive activity and diuretic
activity. In previous studies it has been found that the PGE.sub.2
receptor has various subtypes, each possessing differing
physiological roles. At this time, it is known that the PGE.sub.2
receptor has four primary subtypes denoted EP.sub.1, EP.sub.2,
EP.sub.3 and EP.sub.4, each of which mediates different effects in
various tissues and cells (Coleman, R. A. et al., Pharm. Rev. 1994,
46(2), 205-229). The EP.sub.4 receptor is distributed in such
organs as the thymus, heart, kidney, liver, intestine, womb, ductus
arteriosus and bone, and it is known that the EP.sub.4 receptor is
related to relaxation of smooth muscle, differentiation and
proliferation of lymphocytes, proliferation of mesangial cells, and
collagen production of the fibroblasts. In both the pig and the
dog, modulation of the EP.sub.4 receptor has been characterized
with relaxation of the saphenous vein, and in the rabbit relaxation
of the jugular vein occurs (Coleman, R. A. et al., Prostaglandins
1994, 47, 151).
The EP.sub.4 receptor is also expressed in the ductus arteriosus
(Bhattacharya, M. et al., Circulation 1999, 100, 1751-1756). The
ductus arteriosus is an arterial connection in the fetus, which
directs blood away from the pulmonary circulation and towards the
placenta where oxygenation occurs (Heymann, M. A.; Rudolph, A. M.
Physiol. Rev. 1975, 55, 62-78). In one proposed model the EP.sub.4
receptor in the ductus arteriosus acts as a sensor that responds to
the perinatal drop in circulating levels of PGE.sub.2 by triggering
closure of the ductus arteriosus (Nguyen, M. et al., Nature 1997,
390, 78-81). Closure of the ductus arteriosus was observed in an in
vivo fetal sheep model after administration of a selective EP.sub.4
antagonist (PCT International Application WO 01/42281, published on
Jun. 14, 2001). Maintaining the ductus arteriosus in the open, or
patent state is desirable in the fetus and in infants with certain
types of congenital heart defects where pulmonary or systemic blood
flow depends on patency of the ductus arteriosus. Maintaining
patency of the ductus arteriosus in infants with certain other
types of congenital heart disease such as coarctation of the aorta,
transposition of the great arteries, and Ebstein's anomaly may also
be desirable. For example, infants with coarctation of the aorta, a
condition constituting 7% to 8% of congenital cardiac defects, may
have sudden onset of heart failure, cardiovascular collapse, and
severe metabolic acidosis as the ductus arteriosus closes and
distal perfusion is compromised. In cases such as these, PGE.sub.1
infusions have been utilized to reopen and maintain the patency of
the ductus arteriosus prior to surgical repair of the defect.
An excess of aqueous humor in the anterior chamber of the eye can
result in elevated intraocular pressure or ocular hypertension.
Ocular hypertension is a symptom and/or risk factor for glaucoma, a
disease that can damage the optic nerve and cause blindness. The
EP.sub.4 receptor has been found in ocular tissues involved in the
production of the aqueous humor, such as human ciliary epithelial
cells and human ciliary muscle cells (Mukhopadhyay et al., Biochem.
Pharmacol. 1997, 53, 1249-1255). Trabecular meshwork cells are
known to be involved in the regulation of intraocular pressure
(Clark et al., Investigative Opthalmology & Visual Science
1994, 35, 281-294; and Lutjen-Drescoll, Progress in Retinal and Eye
Research 1998, 18, 91-119). The EP.sub.4 receptor has also been
found in human trabecular meshwork cells and it has been proposed
that activation of the EP.sub.4 receptors in the trabecular
meshwork cells can result in relaxation of these cells, thereby
lowering intraocular pressure (PCT International Patent Application
WO 00/38667, published on Jul. 6, 2000).
As PGE.sub.1 and PGE.sub.2 bind to all four of the PGE.sub.2
receptor subtypes (EP.sub.1, EP.sub.2, EP.sub.3, and EP.sub.4),
various physiological activities may result, some of which may be
an undesired side effect due to the lack of selectivity in binding
to the PGE.sub.2 receptor subtypes. It is therefore desirable to
have methods of treatment for various disorders comprising
administration of compounds with selectivity to a particular
PGE.sub.2 receptor subtype.
Great Britain Patent Specification 1 553 595 discloses compounds of
the formula
##STR00002## wherein the double bonds are cis or trans and the
variables are defined as set forth therein. Those compounds are
disclosed as having spasmogenic and spasmolytic activity, for
example bronchodilatory and antihypertensive effects. The compounds
are also disclosed as having utility in the inhibition of the
secretion of gastric juice and as having abortive effects.
U.S. Pat. No. 4,115,401 discloses compound of the formula
##STR00003## wherein the variables are defined as set forth
therein. Those compounds are disclosed as having spasmogenic,
cardiovascular and bronchodilatory effects.
U.S. Pat. No. 4,113,873 discloses compound of the formula
##STR00004## wherein the variables are defined as set forth
therein. Those compounds are disclosed as having utility as a
bronchodilator, as an antihypertensive agent, as an enhancer of
spontaneous contraction of the uterus and for the treatment of
gastro-intestinal disorders or gastric ulcers.
Great Britain Patent Specification 1 583 163 discloses compounds of
the formula
##STR00005## wherein the variables are defined as set forth
therein. Those compounds are disclosed as having spasmogenic,
bronchodilatory, vasoconstricting, vasodilating and abortive
properties as well as utility in the inhibition of gastric acid
secretion.
U.S. Pat. No. 4,177,346, discloses compounds of the formula
##STR00006## wherein the variables are defined as set forth
therein. Those compounds are disclosed as having vasodilator,
antihypertensive, bronchodilator, antifertility and antisecretory
activity.
U.S. Patent Application Publications Nos. US 2001/0041729, which
published on Nov. 15, 2001, and US 2001/0047105, which published on
Nov. 29, 2001, disclose methods of treatment with compounds of the
formula
##STR00007## wherein the variables are defined as set forth
therein. The methods of treatment disclosed in US 2001/0041729
include the treatment of acute or chronic renal failure or
dysfunction, or a condition caused thereby, such as hypertension,
congestive heart failure, glomerulonephritis, uremia or chronic
renal insufficiency. The methods of treatment disclosed in US
2001/0047105 include the treatment of conditions which present with
low bone mass, particularly osteoporosis, frailty, an osteoporotic
fracture, a bone defect, childhood idiopathic bone loss, alveolar
bone loss, mandibular bone loss, bone fracture, osteotomy, bone
loss associated with periodontitis, or prosthetic ingrowth.
U.S. patent application Ser. No. 09/990,556, which was filed on
Nov. 21, 2001, discloses compounds of the formula
##STR00008## wherein the variables are as defined therein. The
compounds are useful for the treatment of conditions which present
with low bone mass such as osteoporosis, frailty, an osteoporotic
fracture, a bone defect, childhood idiopathic bone loss, alveolar
bone loss, mandibular bone loss, bone fracture, osteotomy, bone
loss associated with periodontis or prosthetic ingrowth.
There exists a continuing need and a continuing search in this
field of art for methods of treating hypertension, liver failure,
loss of patency of the ductus arteriosus, glaucoma and ocular
hypertension. More specifically, there is a need for methods of
treating hypertension, liver failure, loss of patency of the ductus
arteriosus, glaucoma or ocular hypertension in a patient in need
thereof with selective prostaglandin receptor agents that do not
have the undesired side effects caused by methods of treatment with
non-selective agents.
SUMMARY OF THE INVENTION
The present invention is directed to methods of treating
hypertension, liver failure, loss of patency of the ductus
arteriosus, glaucoma or ocular hypertension in a patient,
comprising administering to the patient a selective EP.sub.4
receptor agonist or a prodrug thereof, or a pharmaceutically
acceptable salt of the selective EP.sub.4 receptor agonist or
prodrug.
A first embodiment of the present invention is directed to methods
of treating hypertension, liver failure, loss of patency of ductus
arteriosus, glaucoma or ocular hypertension in a patient,
comprising administering to the patient a therapeutically effective
amount of a selective EP.sub.4 receptor agonist of Formula I
##STR00009## or a prodrug thereof, a pharmaceutically acceptable
salt of the selective EP.sub.4 receptor agonist or prodrug or a
stereoisomer or diastereomeric mixture of the EP.sub.4 receptor
agonist, prodrug or salt, wherein: is a single or double bond; X is
--CH.sub.2-- or O; Z is --(CH.sub.2).sub.3--, thienyl, thiazolyl or
phenyl, provided that when X is O, then Z is phenyl; Q is carboxyl,
(C.sub.1-C.sub.4)alkoxylcarbonyl or tetrazolyl; R.sup.2 is --Ar or
--Ar.sup.1--V--Ar.sup.2; V is a bond, --O--, --OCH.sub.2-- or
--CH.sub.2O--; Ar is a partially saturated, fully saturated or
fully unsaturated five to eight membered ring optionally having one
to four heteroatoms selected independently from oxygen, sulfur and
nitrogen, or a bicyclic ring consisting of two fused independently
partially saturated, fully saturated or fully unsaturated five or
six membered rings, taken independently, optionally having one to
four heteroatoms selected independently from nitrogen, sulfur and
oxygen, said partially or fully saturated ring or bicyclic ring
optionally having one or two oxo groups substituted on carbon or
one or two oxo groups substituted on sulfur; and Ar.sup.1 and
Ar.sup.2 are each independently a partially saturated, fully
saturated or fully unsaturated five to eight membered ring
optionally having one to four heteroatoms selected independently
from oxygen, sulfur and nitrogen, said partially or fully saturated
ring optionally having one or two oxo groups substituted on carbon
or one or two oxo groups substituted on sulfur; said Ar moiety is
optionally substituted on carbon or nitrogen, on one ring if the
moiety is monocyclic, or on one or both rings if the moiety is
bicyclic, with up to three substituents per ring each independently
selected from hydroxy, halo, carboxy, (C.sub.1-C.sub.7)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.7)alkyl, (C.sub.2-C.sub.7)alkenyl,
(C.sub.3-C.sub.7)cycloalkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)alkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)alkanoyl, formyl,
(C.sub.1-C.sub.8)alkanoyl,
(C.sub.1-C.sub.6)alkanoyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.4)alkanoylamino,
(C.sub.1-C.sub.4)alkoxycarbonylamino, hydroxysulfonyl,
aminocarbonylamino or mono-N--, di-N,N--, di-N,N'-- or
tri-N,N,N'--(C.sub.1-C.sub.4)alkyl substituted aminocarbonylamino,
sulfonamido, (C.sub.1-C.sub.4)alkylsulfonamido, amino, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylamino, carbamoyl, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylcarbamoyl, cyano, thiol,
(C.sub.1-C.sub.6)alkylthio, (C.sub.1-C.sub.6)alkylsulfinyl,
(C.sub.1-C.sub.4)alkylsulfonyl and mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylaminosulfinyl, wherein said alkyl and
alkoxy substituents in the definition of Ar are optionally
substituted on carbon with up to three fluoro; and said Ar.sup.1
and Ar.sup.2 moieties are independently optionally substituted on
carbon or nitrogen with up to three substituents each independently
selected from hydroxy, halo, carboxy, (C.sub.1-C.sub.7)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.7)alkyl, (C.sub.2-C.sub.7)alkenyl,
(C.sub.3-C.sub.7)cycloalkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)alkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)alkanoyl, formyl,
(C.sub.1-C.sub.8)alkanoyl,
(C.sub.1-C.sub.6)alkanoyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.4)alkanoylamino,
(C.sub.1-C.sub.4)alkoxycarbonylamino, hydroxysulfonyl,
aminocarbonylamino or mono-N--, di-N,N--, di-N,N'-- or
tri-N,N,N'--(C.sub.1-C.sub.4)alkyl substituted aminocarbonylamino,
sulfonamido, (C.sub.1-C.sub.4)alkylsulfonamido, amino, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylamino, carbamoyl, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylcarbamoyl, cyano, thiol,
(C.sub.1-C.sub.6)alkylthio, (C.sub.1-C.sub.6)alkylsulfinyl,
(C.sub.1-C.sub.4)alkylsulfonyl and mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylaminosulfinyl, wherein said alkyl and
alkoxy substituents in the definition of Ar.sup.1 and Ar.sup.2 are
optionally substituted on carbon with up to three fluoro;
A preferred method of the present invention is a method of the
first embodiment wherein the selective EP.sub.4 receptor agonist is
a compound, designated Group A, of Formula Ia
##STR00010## a prodrug thereof, a pharmaceutically acceptable salt
of said compound or said prodrug, and stereoisomers and
diastereomeric mixtures of said compound, prodrug or salt,
wherein:
##STR00011## and R.sup.2 is Ar wherein said Ar moiety is optionally
substituted on carbon or nitrogen, on one ring if the moiety is
monocyclic, or on one or both rings if the moiety is bicyclic, with
up to three substituents per ring each independently selected from
hydroxy, halo, carboxy, (C.sub.1-C.sub.7)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.7)alkyl, (C.sub.2-C.sub.7)alkenyl,
(C.sub.3-C.sub.7)cycloalkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)alkyl,
(C.sub.3-C.sub.7)cycloalkyl(C.sub.1-C.sub.4)alkanoyl, formyl,
(C.sub.1-C.sub.8)alkanoyl,
(C.sub.1-C.sub.6)alkanoyl(C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.4)alkanoylamino,
(C.sub.1-C.sub.4)alkoxycarbonylamino, hydroxysulfonyl,
aminocarbonylamino or mono-N--, di-N,N--, di-N,N'-- or
tri-N,N,N'--(C.sub.1-C.sub.4)alkyl substituted aminocarbonylamino,
sulfonamido, (C.sub.1-C.sub.4)alkylsulfonamido, amino, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylamino, carbamoyl, mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylcarbamoyl, cyano, thiol,
(C.sub.1-C.sub.6)alkylthio, (C.sub.1-C.sub.6)alkylsulfinyl,
(C.sub.1-C.sub.4)alkylsulfonyl and mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylaminosulfinyl, wherein said alkyl and
alkoxy substituents in the definition of Ar.sup.1 and Ar.sup.2 are
optionally substituted on carbon with up to three fluoro.
Another preferred method of the present invention is a method of
the first embodiment wherein the selective EP.sub.4 receptor
agonist is a compound within Group A, designated Group B, a prodrug
thereof, a pharmaceutically acceptable salt of said compound or
said prodrug, and stereoisomers and diastereomeric mixtures of said
compound, prodrug or salt, wherein Ar is cyclohexyl,
1,3-benzodioxolyl, thienyl, naphthyl or phenyl optionally
substituted with one or two (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl, chloro, fluoro,
trifluoromethyl or cyano, wherein said alkyl and alkoxy
substituents in the definition of Ar are optionally substituted
with up to three fluoro.
Another preferred method of the present invention is a method of
the first embodiment, wherein the selective EP.sub.4 receptor
agonist is a compound within Group B, designated Group C, a prodrug
thereof, a pharmaceutically acceptable salt of said compound or
said prodrug, and stereoisomers and diastereomeric mixtures of said
compound, prodrug or salt, wherein is a single bond; Q is carboxy
or (C.sub.1-C.sub.4)alkoxylcarbonyl; and Z is
##STR00012##
Another preferred method of the present invention is a method of
the first embodiment, in which the selective EP.sub.4 receptor
agonist is a compound within Group C, designated Group D, a prodrug
thereof, a pharmaceutically acceptable salt of said compound or
said prodrug or a stereoisomer or diastereomeric mixture of said
compound, prodrug or salt, wherein Q is carboxy and Ar is phenyl
optionally substituted with one (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl, chloro, fluoro,
trifluoromethyl or cyano, wherein said alkyl and alkoxy
substituents in the definition of Ar are optionally substituted
with up to three fluoro.
Another preferred method of the present invention is a method of
the first embodiment, in which the selective EP.sub.4 receptor
agonist is a compound within Group D, a prodrug thereof, a
pharmaceutically acceptable salt of said compound or said prodrug
or a stereoisomer or diastereomeric mixture of said compound,
prodrug or salt, wherein Ar is 3-trifluoromethylphenyl.
Another preferred method of the present invention is a method of
the first embodiment, in which the selective EP.sub.4 receptor
agonist is a compound within Group D, a prodrug thereof, a
pharmaceutically acceptable salt of said compound or said prodrug
or a stereoisomer or diastereomeric mixture of said compound,
prodrug or salt, wherein Ar is 3-chlorophenyl.
Another preferred method of the present invention is a method of
the first embodiment, in which the selective EP.sub.4 receptor
agonist is a compound within Group D, a prodrug thereof, a
pharmaceutically acceptable salt of said compound or said prodrug
or a stereoisomer or diastereomeric mixture of said compound,
prodrug or salt, wherein Ar is 3-trifluoromethoxyphenyl.
A particularly preferred method of the present invention is a
method of the first embodiment, in which the selective EP.sub.4
receptor agonist is a compound selected from
5-(3-(2S-(3R-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl)-5-oxo-pyrrolidin-
-1-yl)-propyl)-thiophene-2-carboxylic acid;
5-(3-(2S-(3R-hydroxy-4-(3-trifluoromethoxy-phenyl)-butyl)-5-oxo-pyrrolidi-
n-1-yl)-propyl)-thiophene-2-carboxylic acid; or
5-(3-(2S-(4-(3-chloro-phenyl)-3R-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl)-pr-
opyl)-thiophene-2-carboxylic acid.
Another method of the present invention is a method of the first
embodiment, in which the selective EP.sub.4 receptor agonist is a
compound within Group A, a prodrug thereof, a pharmaceutically
acceptable salt of said compound or said prodrug or a stereoisomer
or diastereomeric mixture of said compound, prodrug or salt,
wherein X is --CH.sub.2--, Z is --(CH.sub.2).sub.3--, Q is carboxyl
or (C.sub.1-C.sub.4)alkoxycarbonyl and Ar is phenyl independently
substituted with one to three cyano, (C.sub.1-C.sub.7)alkoxy
substituted with one to three fluoro or
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl.
Another preferred method of the present invention is a method of
the first embodiment in which the selective EP.sub.4 receptor
agonist is a compound within the group of compounds as described in
the immediately preceding paragraph, a prodrug thereof, a
pharmaceutically acceptable salt of said compound or said prodrug,
and stereoisomers and diastereomeric mixtures of said compound,
prodrug or salt, wherein is a single bond; Q is carboxy or
(C.sub.1-C.sub.4)alkoxylcarbonyl; and Z is
##STR00013##
Yet another preferred embodiment of the present invention is a
method of the first embodiment, in which the selective EP.sub.4
receptor agonist is a compound within the group of compounds as
described in the immediately preceding paragraph, a prodrug
thereof, a pharmaceutically acceptable salt of said compound or
said prodrug or a stereoisomer or diastereomeric mixture of said
compound, prodrug or salt, wherein Q is carboxy and Ar is phenyl
optionally substituted with one (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl, chloro, fluoro,
trifluoromethyl or cyano, wherein said alkyl and alkoxy
substituents in the definition of Ar are optionally substituted
with up to three fluoro.
Another preferred embodiment of the present invention is a method
of the first embodiment in which the disorder is hypertension.
Another preferred embodiment of the present invention is a method
of the first embodiment in which the disorder is liver failure. Yet
another embodiment of the present invention is a method of the
first embodiment in which said disorder is loss of patency of the
ductus arteriosus.
Another aspect of the present invention is directed to methods of
treating hypertension, liver failure, loss of patency of the ductus
arteriosus, glaucoma or ocular hypertension in a patient in need
thereof, comprising administering to the patient a pharmaceutical
composition; the pharmaceutical composition comprising a compound
of Formula I or a prodrug thereof, or a pharmaceutically acceptable
salt of the compound or prodrug, or a stereoisomer or
diastereomeric mixture of the compound, prodrug or salt, and a
pharmaceutically acceptable carrier, vehicle or diluent.
Another aspect of the present invention is directed to methods of
treating hypertension with combinations of a compound of Formula I
or a prodrug thereof, or a pharmaceutically acceptable salt of the
compound or prodrug, or a stereoisomer or diastereomeric mixture of
the compound, prodrug or salt; and an HMG-CoA reductase inhibitor
(statin) or a prodrug thereof or a pharmaceutically acceptable salt
of the HMG-CoA reductase inhibitor or prodrug.
Another aspect of the present invention is directed to methods of
treating hypertension with combinations of a compound of Formula I
or a prodrug thereof, or a pharmaceutically acceptable salt of the
compound or prodrug, or a stereoisomer or diastereomeric mixture of
the compound, prodrug or salt; and an antihypertensive agent or a
prodrug thereof or a pharmaceutically acceptable salt of the
antihypertensive agent or prodrug.
Another aspect of the present invention is a kit comprising: a. an
amount of a Formula I compound, a prodrug thereof or a
pharmaceutically acceptable salt of said compound or said prodrug,
or a stereoisomer or diastereomeric mixture of said compound,
prodrug or salt and a pharmaceutically acceptable carrier or
diluent in a first unit dosage form; b. an amount of an
antihypertensive agent, a prodrug thereof, or a pharmaceutically
acceptable salt of said antihypertensive agent or prodrug, and a
pharmaceutically acceptable carrier or diluent in a second unit
dosage form; and c. a container.
Yet another aspect of the present invention is a kit comprising: a.
an amount of a Formula I compound, a prodrug thereof or a
pharmaceutically acceptable salt of said compound or said prodrug,
or a stereoisomer or diastereomeric mixture of said compound,
prodrug or salt and a pharmaceutically acceptable carrier or
diluent in a first unit dosage form; b. an amount of an HMG Co-A
reductase inhibitor, a prodrug thereof, or a pharmaceutically
acceptable salt of said HMG Co-A reductase inhibitor or prodrug,
and a pharmaceutically acceptable carrier or diluent in a second
unit dosage form; and c. a container.
DETAILED DESCRIPTION OF THE INVENTION
The term "treating", "treat" or "treatment" is used herein includes
preventative (e.g. prophylactic), palliative and curative
treatment. The term "therapeutically effective amount", as used
herein, means the amount of selective EP.sub.4 receptor agonist
that will elicit the desired therapeutic effect or provide the
desired benefit when administered according to the desired
treatment regimen. For example, a "therapeutically effective
amount" of a compound of Formula I is an amount that will treat
hypertension, liver failure, loss of patency of the ductus
arteriosus, glaucoma or ocular hypertension in a patient in need
thereof. The term "selective EP.sub.4 receptor agonist" as used
herein means a chemical substance of Formula I that can interact
with the EP.sub.4 receptor and initiate a physiological or
pharmacological response characteristic of the EP.sub.4 receptor
and which has a greater affinity for the EP.sub.4 receptor than for
the EP.sub.1, EP.sub.2 and EP.sub.3 receptors. A preferred group of
selective EP.sub.4 receptor agonists are those compounds of Formula
I that can interact with the EP.sub.4 receptor and initiate a
physiological or pharmacological response characteristic of the
EP.sub.4 receptor and which have approximately a tenfold greater
affinity for the EP.sub.4 receptor than for the EP.sub.1, EP.sub.2
and EP.sub.3 receptors. The term "loss of patency of the ductus
arteriosus" as used herein means the partial or complete closure of
the ductus arteriosus. The term "pharmaceutically acceptable" as
used herein means that the carrier, vehicle, diluent, excipients,
and/or salt must be compatible with the other ingredients of the
formulation, and not deleterious to the recipient thereof. The term
"prodrug" refers to compounds that are drug precursors which,
following administration, release the drug in vivo by some chemical
or physiological process (e.g. a prodrug on being brought to
physiological pH or through enzyme action is converted to the
desired drug form). Exemplary prodrugs upon cleavage release the
corresponding drug compound.
The term "hydroxy" as used herein means the group --OH. The term
"thiol" as used herein means the group --SH. The term "cyano" as
used herein means the group --CN. The term "halo" as used herein
means fluoro, chloro, bromo and iodo. The term "carboxy" as used
herein means the group --CO.sub.2H. The term "carbonyl" as used
herein means the group --C(O)--. The term "formyl" as used herein
means the group --C(O)H. The term "amino" means the group
--NH.sub.2, except when the amino group is mono or disubstituted,
in which case one or both of the --NH.sub.2 hydrogens is
substituted as specified. The term "(C.sub.1-C.sub.7)alkyl" as used
herein means a straight or branched chain hydrocarbon group having
from one to seven carbons. The term "(C.sub.1-C.sub.7)alkyl"
includes, but is not limited to, groups such as methyl, ethyl,
propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl,
neopentyl, methylpentyl, hexyl, heptyl, methylhexyl and the like.
Likewise, other alkyl terms such as "(C.sub.1-C.sub.4)alkyl",
"(C.sub.1-C.sub.6)alkyl" and "(C.sub.1-C.sub.8)alkyl" are straight
or branched chain hydrocarbon groups with one to four, one to six,
and one to eight carbons, respectively. The term
"(C.sub.2-C.sub.7)alkenyl" means a straight or branched chain
hydrocarbon group having two to seven carbons and a carbon-carbon
double bond. The term "(C.sub.2-C.sub.7)alkenyl" includes, but is
not limited to, groups such as vinyl, propenyl, allyl,
2-methylpropenyl, butenyl, etc. The term
"(C.sub.3-C.sub.7)cycloalkyl" as used herein means a cyclic
hydrocarbon group having from three to seven carbons. The term
"(C.sub.3-C.sub.7)cycloalkyl" includes, but is not limited to,
groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, methylcyclopropyl, ethylcyclopropyl, methylcyclobutyl,
etc. The terms "(C.sub.1-C.sub.7)alkoxy" and
"(C.sub.1-C.sub.4)alkoxy", as used herein, mean the groups
(C.sub.1-C.sub.7)alkyl-O-- and (C.sub.1-C.sub.4)alkyl-O--,
respectively. For example, the term "(C.sub.1-C.sub.4)alkoxy"
includes methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy,
sec-butoxy and tert-butoxy. The terms "(C.sub.1-C.sub.8)alkanoyl",
"(C.sub.1-C.sub.6)alkanoyl", and "(C.sub.1-C.sub.4)alkanoyl", as
used herein, mean the groups (C.sub.1-C.sub.8)alkyl-C(O)--,
(C.sub.1-C.sub.6)alkyl-C(O)--, and (C.sub.1-C.sub.4)alkyl-C(O)--,
respectively. The term "(C.sub.1-C.sub.4)alkanoylamino as used
herein means the group (C.sub.1-C.sub.4)alkyl-C(O)NH--. The term
"(C.sub.1-C.sub.4)alkoxycarbonylamino" as used herein means the
group (C.sub.1-C.sub.4)alkyl-O--C(O)--NH--. The term
"hydroxysulfonyl" as used herein means the group --SO.sub.3H. The
term "aminocarbonylamino" as used herein means the group
--NHC(O)NH.sub.2. The terms "mono-N--, di-N,N--, di-N,N'--, or
tri-N,N,N'--(C.sub.1-C.sub.4)alkyl substituted aminocarbonylamino",
as used herein, mean the groups --NHC(O)NH(C.sub.1-C.sub.4)alkyl,
--NHC(O)N((C.sub.1-C.sub.4)alkyl).sub.2,
--N((C.sub.1-C.sub.4)alkyl)C(O)NH(C.sub.1-C.sub.4)alkyl, or
--N((C.sub.1-C.sub.4)alkyl)C(O)NH((C.sub.1-C.sub.4)alkyl).sub.2,
respectively. The term "sulfonamido" as used herein means the group
--S(O).sub.2NH.sub.2. The terms mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylamino as used herein mean the groups
--NH(C.sub.1-C.sub.4)alkyl or --N((C.sub.1-C.sub.4)alkyl).sub.2,
respectively. The term "carbamoyl" as used herein means the group
--OC(O)NH.sub.2. The terms "mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylcarbamoyl" mean the groups
--OC(O)NH(C.sub.1-C.sub.4)alkyl or
--OC(O)N((C.sub.1-C.sub.4)alkyl).sub.2, respectively. The term
"(C.sub.1-C.sub.6)alkylthio" as used herein means the group
(C.sub.1-C.sub.6)alkyl-S--. The term
"(C.sub.1-C.sub.6)alkylsulfinyl" as used herein means the group
(C.sub.1-C.sub.6)alkyl-S(O)--. The term
"(C.sub.1-C.sub.4)alkylsulfonyl" as used herein means the group
(C.sub.1-C.sub.4)alkyl-S(O).sub.2--. The terms "mono-N-- or
di-N,N--(C.sub.1-C.sub.4)alkylaminosulfinyl" as used herein mean
the groups --S(O)NH(C.sub.1-C.sub.4)alkyl or
--S(O)N((C.sub.1-C.sub.4)alkyl).sub.2, respectively.
The term "pharmaceutically acceptable salt" as used herein refers
to both nontoxic anionic salts and cationic salts. Anionic salts
include, but are not limited to, chloride, bromide, iodide,
sulfate, bisulfate, phosphate, acetate, maleate, fumarate, oxalate,
lactate, tartrate, citrate, gluconate, methanesulfonate and
4-toluene sulfonate. Cationic salts include, but are not limited
to, sodium, potassium, calcium, magnesium, ammonium, protonated
benzathine (N,N'-dibenzylethylenediamine), choline, ethanolamine,
diethanolamine, ethylenediamine, meglamine (N-methyl glucamine),
benethamine (N-benzylphenethylamine), piperazine or tromethamine
(2-amino-2-hydroxymethyl-1,3-propanediol.
The chemist of ordinary skill in the art will also recognize that
certain compounds of Formula I of this invention can exist in
tautomeric form, i.e., that a rapid equilibrium exists between two
isomers. A common example of tautomerism is keto-enol tautomerism,
i.e.,
##STR00014## Examples of compounds that can exist as tautomers
include hydroxypyridines, hydroxypyrimidines and hydroxyquinolines.
Other examples of compounds that can exist as tautomers will be
recognized by those skilled in the art. All such tautomers and
mixtures thereof are included in this invention.
The methods of the present invention also includes the use of
isotopically-labeled compounds, which are identical to those
recited in Formula I, but for the fact that one or more atoms are
replaced by an atom having an atomic mass or mass number different
from the atomic mass or mass number usually found in nature.
Examples of isotopes that can be incorporated into compounds of
Formula I include isotopes of hydrogen, carbon, nitrogen, oxygen,
phosphorous, sulfur, fluorine and chlorine, such as .sup.2H,
.sup.3H, .sup.13C, .sup.14C, .sup.15N, .sup.18O, .sup.17O,
.sup.31P, .sup.32P, .sup.35S, .sup.18F and .sup.36Cl, respectively.
Methods of treatment with compounds of Formula I, prodrugs thereof,
and pharmaceutically acceptable salts of said compounds and said
prodrugs, and stereoisomers and diastereomeric mixtures of said
compounds, prodrugs and salts, which contain the aforementioned
isotopes and/or other isotopes of other atoms are within the scope
of this invention. Certain isotopically-labeled compounds of
Formula I, for example those into which radioactive isotopes such
as .sup.3H and .sup.14C are incorporated, are useful in drug and/or
substrate tissue distribution assays. Tritiated, i.e., .sup.3H, and
carbon-14, i.e., .sup.14C, isotopes are particularly preferred for
their ease of preparation and detectability. Further, substitution
with heavier isotopes such as deuterium, i.e., .sup.2H, can afford
certain therapeutic advantages resulting from greater metabolic
stability, for example increased in vivo half-life or reduced
dosage requirements and, hence, may be preferred in some
circumstances. Isotopically labeled compounds of Formula I and
prodrugs thereof can generally be prepared by carrying out the
procedures disclosed in the Schemes and/or as described for the
Compounds and Preparations below, by substituting a readily
available isotopically labeled reagent for a non-isotopically
labeled reagent.
The compounds of Formula I used in the methods of this invention
have asymmetric carbon atoms, and therefore, are enantiomers or
diastereomers. Diasteromeric mixtures can be separated into their
individual diastereomers on the basis of their physical chemical
differences by methods known per se, for example, by chromatography
and/or fractional crystallization. Enantiomers can be separated by
converting the enantiomeric mixture into a diasteromeric mixture by
reaction with an appropriate optically active compound (e.g.,
alcohol), separating the diastereomers and converting (e.g.,
hydrolyzing) the individual diastereomers to the corresponding pure
enantiomers. Enantiomers and diastereomers of the compounds of
Formula I can also be prepared by utilizing suitable
enantiomerically enriched starting materials, or by asymmetric or
diastereoselective reactions to introduce asymmetric carbon atoms
with the correct stereochemistry. All such isomers, including
diastereomers, enantiomers and mixtures thereof are considered as
compounds of Formula I and can be used in the methods of this
invention. Some of the compounds of Formula I are acidic, and
therefore, can form a salt with a pharmaceutically acceptable
cation. All such salts are within the scope of the compounds of
Formula I and can be prepared by conventional methods. For example,
the salt can be prepared simply by contacting the acidic and basic
entities, usually in a stoichiometric ratio, in either an aqueous,
non-aqueous or partially aqueous medium, as appropriate. The salts
are recovered either by filtration, by precipitation with a
non-solvent followed by filtration, by evaporation of the solvent,
or, in the case of aqueous solutions, by lyophilization, as
appropriate.
The present invention is directed to the treatment of disorders
responsive to modulation of the EP.sub.4 receptor by administering
to a patient in need thereof a therapeutically effective amount of
a compound of Formula I. More specifically, the present invention
is directed to the treatment of hypertension, liver failure, loss
of patency of ductus arteriosus, glaucoma or ocular hypertension by
administration of a selective EP.sub.4 receptor agonist of Formula
I. The compounds of Formula I, which are useful in the methods of
the present invention, are prepared as described in U.S. patent
application Ser. No. 09/990,556, which was filed on Nov. 21, 2001.
In general, the compounds of Formula I are made by processes that
are analogous to those known in the chemical arts. These processes
include methods that may require protection of remote functionality
(e.g., primary amine, secondary amine, secondary alcohol, primary
alcohol, carboxyl in Formula I precursors). The need for such
protection will vary depending upon the nature of the remote
functionality and the conditions of the preparation methods. The
need for such protection is readily determined by one skilled in
the art. The use of such protection/deprotection methods is also
within the skill in the art. The term "protecting group," where
used herein, refers to a radical that may be attached to a
functional group on a substrate. The "protecting group" is such
that it is easily attached and easily removed without affecting
other functional groups of the substrate and it prevents the
protected functional group from being removed, altered or otherwise
destroyed. For a general description of protecting groups and their
use, see Greene, T. W.; Wuts, P. G. M., Protective Groups in
Organic Synthesis, 2.sup.nd ed.; John Wiley and Sons Inc.: New
York, 1991. The starting materials and reagents used for the
synthesis of compounds of Formula I are also readily available or
can be easily synthesized by those skilled in the art using
conventional methods of organic synthesis in light of this
disclosure.
In general, compounds of Formula I are prepared by protection of
the hydroxyl group of either racemic or
(R)-hydroxymethyl-2-pyrrolidinone, followed by alkylation of the
amide nitrogen with an alkyl halide that contains a suitably
protected acid precursor or isostere (Scheme A). The term
"isostere," where used herein, refers to a functional group that,
when used in place of another functional group, approximates the
reactivity of the functional group that it replaces. In some cases,
the alkyl halide must be further elaborated to install the suitably
protected acid precursor or isostere (Scheme B1). The hydroxyl
protecting group is removed, the alcohol oxidized to the aldehyde
which is then reacted with the anion of a suitable keto-phosphonate
(Scheme C). The resulting enone of Formula 8 of Scheme E is then
subjected to reduction of both the double bond and ketone to give
the desired saturated alcohols of Formula 9 of Scheme E. If
desired, a diastereoselective reduction of the enone can be
effected to give, for example, predominantly the 15-(R) isomer or
the 15-(S) isomer. The carboxylic ester or precursor to an acid
isostere (e.g., nitrile) is then converted into the appropriate
acidic group (carboxylic acid, tetrazole, etc).
A preferred method for converting a nitrile into the desired
tetrazole is treatment of the nitrile with dibutyltin oxide and
trimethylsilylazide, in refluxing toluene (S. J. Wittenberger and
B. G. Donner, J. Org. Chem. 1993, 58, 4139-4141). For a review of
alternative preparations of tetrazoles see R. N. Butler,
Tetrazoles, in Comprehensive Heterocyclic Chemistry; Potts, K. T.
Ed.; Pergamon Press: Oxford, 1984, Vol. 5, pp 791-838.
##STR00015## More specifically, compounds of Formula I are prepared
by the following procedures. In the first general sequence, which
begins with Scheme A, the hydroxyl group of
5-(R)-hydroxymethyl-2-pyrrolidinone (Aldrich Chemical, or prepared
as described by Bruckner et al., Acta. Chim. Hung. Tomus, 1959, 21,
106) is suitably protected (where PG is a suitable protecting
group) by reaction of a compound of Formula 1 in a reaction inert
solvent. As used herein, the expressions "reaction inert solvent"
and "inert solvent" refer to a solvent or mixture of solvents that
does not interact with starting materials, reagents, intermediates
or products in a manner that adversely affects the yield of the
desired product. In some cases herein, a list of preferred reaction
inert solvents, is described. However, any solvent that meets the
above definition of reaction inert solvent for a particular
reaction may be used in that reaction. All reactions are carried
out in a reaction inert solvent unless specifically stated
otherwise. Any standard alcohol protecting group may be utilized,
including tetrahydropyranyl, trimethylsilyl,
tert-butyl-dimethylsilyl, or benzyl. A preferred protecting group
is tert-butyl-dimethylsilyl (TBS), which can be installed by
standard methods as described in Greene, T. W.; Wuts, P. G. M.,
Protective Groups in Organic Synthesis, 2.sup.nd ed.; John Wiley
and Sons Inc.: New York, 1991. It is preferable to treat
5-(R)-hydroxymethyl-2-pyrrolidinone in methylene chloride at
0.degree. C. with 0.1 equivalents (eq.) of 4-dimethylaminopyridine,
1.1 eq. of tert-butyl-dimethylsilylchloride, and 2 eq. of imidazole
(see, e.g., Tetrahedron Asymmetry 1996, 7, 2113). The amide
nitrogen is alkylated with one of a variety of alkylating agents
(hal-CH.sub.2CH.sub.2--X-Z-QP, where hal is a leaving group such as
bromide or iodide, X and Z are as described in the Summary, and QP
is a nitrile, carboxylic acid ester or other precursor to a
carboxylic acid or acid isostere) to introduce the desired side
chain. The amide nitrogen is first deprotonated with a suitable
base. Preferred bases include sodium hexamethyldisilazide (also
referred to herein as NaHMDS or NaN(SiMe.sub.3).sub.2) or sodium
hydride in a reaction inert solvent such as N,N-dimethylformamide
(DMF), tetrahydrofuran (THF), 1,2-dimethoxyethane or 1,4-dioxane. A
preferred solvent is DMF. The appropriate temperature range for
anion formation is between -78.degree. C. and the temperature at
which the solvent refluxes. A preferred temperature for this
reaction is about 0.degree. C. After formation of the anion, the
alkylating agent (hal-CH.sub.2CH.sub.2--X-Z-QP) is added and the
solution is stirred at an appropriate temperature. The appropriate
temperature range for alkylation is between -20.degree. C. and the
temperature at which the solvent refluxes. The preferred
temperature range for this reaction is between 0.degree. C. and
100.degree. C. Typical alkylating agents are primary, secondary,
benzylic, propargyllic halides and primary, secondary, benzylic or
propargyllic sulfonates. Preferred alkylating agents are alkyl
bromides or alkyl iodides.
Many of the useful alkylating agents of the formula
hal-CH.sub.2CH.sub.2--X-Z-QP are commercially available. For
example, ethyl-7-bromoheptanoate and 7-bromoheptanonitrile may be
obtained from Aldrich Chemical, Milwaukee, Wis. Numerous methods
known to those skilled in the art exist for the synthesis of those
and other desired alkylating agents used in the above Scheme (see,
e.g., "The Chemistry of the Carbon-Halogen Bond," Ed. S. Patai, J.
Wiley, New York, 1973 and/or "The Chemistry of Halides,
Pseudo-Halides, and Azides," Eds. S. Patai and Z. Rappaport, J.
Wiley, New York, 1983).
Alkyl halides are also prepared by halogenation of an alcohol or an
alcohol derivative. Alkyl chlorides are typically prepared from the
alcohols with reagents such as hydrogen chloride, thionyl chloride,
phosphorous pentachloride, phosphorous oxychloride or
triphenylphosphine/carbon tetrachloride in a reaction inert
solvent. For the preparation of alkyl bromides the alcohol is
commonly treated with reagents such as hydrogen bromide,
phosphorous tribromide, triphenylphosphine/bromine or
carbonyldiimidazole/allyl bromide in a reaction inert solvent. To
prepare alkyl iodides, the alcohol is typically reacted with
reagents such as triphenylphosphine/iodine/imidazole or hydrogen
iodide in a reaction inert solvent. Alkyl chlorides are converted
to the more reactive alkyl bromides or alkyl iodides by treatment
with an inorganic salt such as sodium bromide, lithium bromide,
sodium iodide or potassium iodide in a reaction inert solvent such
as acetone or methyl ethyl ketone. Alkyl sulfonates are also used
as electrophiles or are converted to alkyl halides. Sulfonates are
prepared from the alcohol using a mild base such as triethylamine
or pyridine and a sulfonyl chloride in a reaction inert solvent
such a methylene chloride or diethyl ether. Conversion to the
halide is accomplished by treatment of the alkyl sulfonate with an
inorganic halide (sodium iodide, sodium bromide, potassium iodide,
potassium bromide, lithium chloride, lithium bromide, etc) or a
tetrabutylammonium halide in a reaction inert solvent.
Alkyl halides of the formula hal-CH.sub.2CH.sub.2--X-Z-QP where X
is CH.sub.2 and Z is phenyl, thienyl or thiazolyl are also prepared
as shown in Scheme B1. For example, propargyl alcohol is treated
with a compound of formula 14 of Scheme B1 containing the suitably
protected acid isostere (hal-Z-QP), where the "hal-Z" group is an
aryl bromide, iodide or triflate, in the presence of copper (I)
iodide; a palladium catalyst such as palladium chloride,
bis(triphenylphosphine)palladium dichloride or
tetrakis(triphenylphosphine)palladium(0); and an amine such as
triethylamine, diisopropylamine or butylamine in a reaction inert
solvent, preferably an aprotic solvent such as acetonitrile, at a
temperature of about 0.degree. C. to about 100.degree. C. For
additional references, see Tetrahedron 1984, 40, 1433 and Org.
Lett. 2000, 2(12), 1729. The resulting alkynes are then converted
to the corresponding alkanes via hydrogenation in the presence of a
palladium or platinum catalyst in a reaction inert solvent such as
methanol, ethanol and/or ethyl acetate at a temperature of about
0.degree. C. to about 50.degree. C. The alcohol portion of the
molecule is replaced with a suitable leaving group such as bromide
or iodide. For the preparation of alkyl bromides, the alcohol is
commonly treated with reagents such as hydrogen bromide,
phosphorous tribromide, triphenylphosphine/bromine or
carbonyldiimidazole/allyl bromide. The use of
carbonyldiimidazole/allyl bromide is preferred. To prepare alkyl
iodides, the alcohol is typically reacted with a reagent such as
triphenylphosphine/iodine/imidazole or hydrogen iodide in a
reaction inert solvent. Alkyl chlorides are converted to the more
reactive alkyl bromides or alkyl iodides by treatment with an
inorganic salt such as sodium bromide, lithium bromide, sodium
iodide or potassium iodide in a reaction inert solvent such as
acetone or methyl ethyl ketone. Alkyl sulfonates can be used as
electrophiles or are converted to alkyl halides. Alkyl sulfonates
are prepared from the corresponding alcohol using a mild base such
as triethylamine or pyridine and a sulfonyl chloride in a reaction
inert solvent such as methylene chloride or diethyl ether.
Conversion to the halide is accomplished by treating the alkyl
sulfonate with an inorganic halide such as, for example, sodium
iodide, sodium bromide, potassium iodide, potassium bromide,
lithium chloride or lithium bromide in a reaction inert solvent.
Conversion to the halide may also be accomplished by treating the
alkyl sulfonate with an organic ammonium halide such as
tetrabutylammonium halide in a reaction inert solvent. Alkyl
chlorides are typically prepared from the alcohols with reagents
such as hydrogen chloride, thionyl chloride, phosphorous
pentachloride, phosphorous oxychloride, or
triphenylphosphine/carbon tetrachloride.
##STR00016##
In some cases, as shown in Scheme B2, it is preferred to first
alkylate with propargyl bromide or iodide, and then further
elaborate to introduce the suitably protected acid precursor or
isostere. For example, where the alkylating agent is propargyl
bromide or iodide, compounds of Formula 3 of Scheme B2 are treated
with compounds of Formula 14 of Scheme B2 containing the suitably
protected acid precursor or isostere (hal-Z-QP), where the "hal-Z"
group is an aryl bromide, iodide or triflate, in the presence of
copper (I) iodide; a palladium catalyst such as palladium chloride,
bis(triphenylphosphine)palladium dichloride or
tetrakis(triphenylphosphine)palladium(0); and an amine such as
triethylamine, diisopropylamine or butylamine in a reaction inert
solvent, preferably an aprotic solvent such as acetonitrile, at a
temperature of about 0.degree. C. to about 100.degree. C. For
additional references see Tetrahedron 1984, 40, 1433 and Org. Lett.
2000, 2(12), 1729. The resulting alkynes are then converted to the
corresponding alkanes via hydrogenation in the presence of a
palladium or platinum catalyst in a reaction inert solvent such as
methanol, ethanol and/or ethyl acetate at a temperature of about
0.degree. C. to about 50.degree. C.
##STR00017##
Halo-arylesters and halo-aryinitriles of Formula 14 of Scheme B2
are prepared by methods known to those skilled in the art. For
example, 2-bromo-4-(ethoxycarbonyl)thiazole is prepared according
to the procedure described in J. Org. Chem. 1996, 61(14), 4623; and
2-bromo-5-(ethoxycarbonyl)thiazole is prepared according to the
procedure described in Helv. Chim. Acta 1942, 25, 1073. Other
halo-arylesters and halo-arylnitriles of Formula 14 of Scheme B2,
which are useful in the procedures of this invention, such as,
inter alia, ethyl-4-bromobenzoate and 4-bromobenzonitrile are
commercially available. Ethyl-2-bromo-thiophene-5-carboxylate is
prepared by esterification of commercially available
2-bromo-thiophene-5-carboxylic acid.
The alcohol protecting groups of compounds of Formula 2 of Scheme A
or Formula 4 of Scheme B2 are then removed. For a general
description of methods for deprotection of protected alcohols, see
Greene, T. W.; Wuts, P. G. M., Protective Groups in Organic
Synthesis, 2.sup.nd ed.; John Wiley and Sons Inc.: New York, 1991.
Removal of the tert-butyl-dimethylsilyl group in compounds of
Formula 2 and Formula 4 of Scheme B2 is preferably accomplished by
treating the compound with tetrabutylammonium fluoride or
trifluoroacetic acid in a reaction inert solvent, preferably in a
suitable aprotic solvent at a temperature of about of -30.degree.
C. to about ambient temperature. Where used herein, the term
"ambient temperature" refers to the temperature of the immediate,
unaltered surroundings of the reaction mixture. Ambient temperature
is generally between 20.degree. C. and 25.degree. C. An especially
preferred solvent is methylene chloride. A preferred temperature
range is between 0.degree. C. to ambient temperature. Another
preferred method to remove the TBS group is by treatment of the
silyl ether with an aqueous solution of a mineral acid in a protic
solvent. In this case, it is preferred that the silyl ether is
treated with a 1N aqueous solution of hydrochloric acid in methanol
at ambient temperature. Subsequent to deprotection, the alcohols
are oxidized to the aldehyde by use of a modification of the
Pfitzner Moffatt oxidation [K. E. Pfitzner and M. E. Moffatt, J.
Am. Chem. Soc. 1965, 87, 5661] which minimizes racemization by
avoiding contact with water. For example, oxidation of the alcohol
to the aldehyde is achieved by stirring the alcohol in a reaction
inert solvent, preferably a hydrocarbon solvent such as toluene,
xylene or, preferably, benzene, with dimethyl sulfoxide, a weak
acid such as acetic acid or, preferably, pyridinium
trifluoroacetate, and a diimide such as diethyl carbodiimide or,
preferably, dimethylaminopropylethylcarbodiimide or, if desired,
dimethylaminopropylethylcarbodiimide hydrochloride, at temperatures
of about 0.degree. C. to about ambient temperature for about one to
about four hours. Alternate methods to achieve oxidation while
minimizing racemization of the asymmetric center adjacent to the
resulting aldehyde are discussed in detail in Tetrahedron Letters
2000, 41, 1359, and include the usual Pfitzner-Moffatt reaction,
oxidation with chromium trioxide-pyridine complex [J. Org. Chem.
1970, 35, 4000], oxidation with Dess-Martin reagent [J. Org. Chem.
1983, 48, 4155] or oxidation with TEMPO-bleach [Tetrahedron Letters
1992, 33, 5029].
The resulting aldehyde is preferably subjected without purification
to a Horner-Wittig reaction with the sodium or lithium salt of a
phosphonate of Formula 7 of Scheme C (R is lower alkyl, haloalkyl
or aryl). The sodium or lithium salts are pre-formed by prior
treatment of the phosphonates with a suitable base such as sodium
hydride or NaN(SiMe.sub.3).sub.2 in a suitable reaction inert
solvent, preferably an aprotic ethereal solvent at a temperature of
about 0.degree. C. to about 50.degree. C. A preferred solvent is
THF and a preferred temperature is ambient temperature. A solution
of the aldehyde is then added to the salt of the phosphonate in a
reaction inert solvent, preferably an aprotic solvent at a
temperature of about 0.degree. C. to about 50.degree. C. to give
enones of Formula 8 of Scheme C. A preferred solvent is THF and a
preferred temperature is ambient temperature.
##STR00018##
Methods for the preparation of phosphonates of Formula 7 of Scheme
C1 can be found in U.S. Pat. No. 3,932,389; U.S. Pat. No.
4,177,346; Tetrahedron Lett. 1989, 30(36), 4787-4790; and Angew.
Chem. 1996, 108(3), 366-369. In general, as shown in Scheme C1, the
phosphonates of Formula 7 are prepared from reaction of the
appropriately substituted arylacetic acid esters or the
methoxymethyl amide of the arylacetic acid with the lithium reagent
derived from a dialkyl methylphosphonate. These methods are also
applicable to cycloalkylacetic esters and methoxymethylamides such
as ethyl-cyclohexylacetate and ethyl-cyclopentylacetate. The aryl-
and cycloalkyl-acetic acid esters are prepared by esterification of
the corresponding acetic acid by methods known to those skilled in
the art. The methoxymethylamides are prepared by a standard amide
bond forming reaction between the corresponding acetic acid and
methoxymethyl amine. Preferably, the coupling of the amine with the
carboxylic acid is carried out in a reaction inert solvent such as
dichloromethane or DMF by a coupling reagent such as
1-(3-dimethylaminopropyl-3-ethylcarbodiimide hydrochloride (EDC) or
1,3-dicyclohexylcarbodiimide (DCC) in the presence of an acid
activating agent such as 1-hydroxybenzotriazole hydrate (HOBT) to
generate the methoxymethyl amide. In the case where the amine is
present as the hydrochloride salt, it is preferable to add one
equivalent of a suitable base such as triethylamine to the reaction
mixture. Alternatively, coupling of the amine with the carboxylic
acid is effected with a coupling reagent such as
benzotriazol-1-yloxy-tris(dimethylamino)-phosphonium
hexafluorophosphate (BOP) in a reaction inert solvent such as
methanol. Such coupling reactions are generally conducted at
temperatures of about -30.degree. C. to about 80.degree. C.,
preferably about 0.degree. C. to about 25.degree. C. For a
discussion of other conditions used for amide couplings, see
HeubenWeyl, Vol. XV, part 11, E. Wunsch, Ed., George Theime Verlag,
1974, Stuttgart.
##STR00019##
The requisite arylacetic acids and esters of Formula 6 of Scheme C1
are commercially available or are prepared by methods well known to
those skilled in the art. As shown in Scheme C2, many aryl and
heteroaryl substituted aryl acetic acids are prepared by Suzuki
couplings of the appropriate arylboronic acids or arylboronate
esters with the desired aryl halides (for a review of the Suzuki
coupling reaction see A. R. Martin and Y. Yang in Acta Chem. Scand.
1993, 47, 221 or J. Am. Chem. Soc. 2000, 122(17), 4020). For
example, the 3-pinacolboronate ester of ethyl-3-bromophenylacetate
is prepared using the method described by Masuda et al. in J. Org.
Chem. 2000, 65, 164. The 3-pinacolboronate ester of
ethyl-3-bromophenylacetate is then coupled with the desired aryl
halide to give the desired 3-aryl-phenylacetic acid (see Synlett.
2000, 6, 829). Hydroxy substituted aryl acetic esters are alkylated
with alkyl halides and benzylic halides by methods well known to
those skilled in the art.
##STR00020##
For a review of the preparation of diaryl ethers, see Angew. Chem.
Int. Ed. 1999, 38(16), 2345. Aryl acetic acids substituted with an
alkylether linkage are prepared using Mitsunobu conditions (for a
review see Synthesis 1981, 1). Typically, the coupling between a
phenolic component and a benzylic alcohol is achieved by addition
of triphenylphosphine and diethyl azodicarboxylate or diisopropyl
azodicarboxylate in a reaction inert solvent such as methylene
chloride or THF.
Alternatively, phosphonates of Formula 7 of Scheme D are prepared
as shown in Scheme D. In general, triethylphosphite is added slowly
to epibromo- or epichloro-hydrin (10) at a temperature of about
135.degree. C. As the triethylphosphite is added, the temperature
drops to about 105.degree. C. The reaction mixture is refluxed
overnight and the product, a compound of Formula 11, is isolated by
vacuum distillation (see Phosphorus, Sulfur Silicon Relat. Elem.
1992, 165, 71, or U.S. Pat. No. 2,627,521). The required Grignard
solutions are prepared from the appropriate aryl halides according
to procedures well known to those skilled in the art in a reaction
inert solvent, preferably an ethereal solvent such as THF, and
cooled to approximately -30.degree. C. Catalytic copper (I) iodide
is added followed by addition of the epoxide of Formula 11
[Phosphorus, Sulfur Silicon Relat. Elem. 1995, 105, 45]. The
requisite aryl halides (e.g., 3-bromo-biphenyl) are commercially
available or are prepared by methods well known to those skilled in
the art.
The resulting alcohols are then oxidized, preferably using a Swern
oxidation [Synthesis 1981,165-185] or Dess-Martin reagent [J. Org.
Chem. 1983, 48, 4155]. Alternative oxidation procedures such as
Pfitzner-Moffatt reaction, chromium trioxide-pyridine complex [R.
Ratcliffe, et al., J. Org. Chem. 1970, 35, 4000], TEMPO-bleach
[Tet. Lett. 1992, 33, 5029], Jones oxidation, Manganese dioxide,
pyridiniumchlorochromate or pyridinium dichromate may also be
utilized to prepare keto-phosphonates of Formula 7 of Scheme D.
##STR00021##
An enone of Formula 8 of Scheme E (which may also be prepared as
shown in Scheme C) is reduced to a mixture of alcohol diastereomers
of Formula 9 of Scheme E by methods well known to those skilled in
the art. In general, the double bond of the enone is first reduced
by catalytic hydrogenation. It is preferred that the double bond is
reduced by hydrogenation over a noble metal catalyst such as
palladium on carbon or platinum oxide in a reaction inert solvent
such as ethyl acetate, methanol or ethanol at a temperature of
ambient temperature up to about the reflux temperature of the
solvent being used under 1 to 4 atmospheres of hydrogen. The
resulting ketone is then treated with a reducing agent, preferably
sodium borohydride, in a protic solvent, preferably ethanol or
methanol, to give alcohols of Formula 9 of Scheme E. Other
selective reducing agents well known to those skilled in the art
that will reduce the ketone but no other functional groups, such as
zinc borohydride or lithium triethylborohydride may also be
employed. The temperature selection will be based upon the activity
of the reducing agent and will preferably be between about
0.degree. C. to ambient temperature. If desired, the mixture of
alcohols of Formula 9 may be separated by preparative
chromatography or HPLC to give the desired 15-(R) diastereomer.
In the second sequence shown in Scheme E, an enone of Formula 8 is
first treated with a hydride reducing agent in the presence of a
chiral catalyst. Where used herein, the term "hydride reducing
agent" refers to a compound that is able to reduce a compound
having a higher oxidation state by transferring hydrogen to the
higher oxidation state compound. A preferred hydride reducing agent
is catecholborane. A preferred chiral catalyst for performing such
reactions enantioselectively is (R)-2-methyl-CBS-oxazaborolidine
reagent (Aldrich Chemical Co., Milwaukee, Wis.) (see the method
described in Eur. J. Org. Chem. 1999, 2655). The reduction is
carried out in a reaction inert solvent, preferably an aprotic
solvent such as methylene chloride, at a temperature of about
-100.degree. C. to ambient temperature. A preferred temperature for
this reaction is about -40.degree. C. Alternative methods and
catalysts which are utilized to effect stereoselective reduction of
the enone carbonyl are described in J. Am. Chem. Soc. 1995, 117,
2675; J. Am. Chem. Soc. 1979, 101, 5843; Tett. Lett. 1990, 31, 611;
U.S. Pat. No. 6,037,505; and Angew. Chem. Int. Ed. 1998, 37, 1986.
The double bond of the allylic alcohol is then reduced to provide
the compound of Formula 9a. It is preferred that the double bond is
reduced by hydrogenation over a noble metal catalyst such as
palladium on carbon or platinum oxide in a reaction inert solvent
such as ethyl acetate, methanol or ethanol at ambient temperature
to the reflux temperature of the solvent being used under 1 to 4
atmospheres of hydrogen.
##STR00022##
A procedure for the preparation of compounds of formula 9b is shown
in Scheme F. In general, tetrahydro-pyrrolizine-3,5-dione (the
compound of formula 12 of Scheme F) is prepared as described in
U.S. Pat. No. 4,663,464 or J. Med. Chem. 1987, 30(3); 498-503. The
compound of Formula 12 of Scheme F is then dissolved in a reaction
inert solvent, preferably an aprotic solvent at a suitable
temperature. It is preferred that said compound is dissolved in
methylene chloride at about 0.degree. C. The reaction mixture is
then treated with the appropriate Grignard reagent (for additional
references on addition of Grignard reagents to Formula 12 of Scheme
F, see Syn. Comm. 1988, 18(1), 37-44; Helv. Chim. Acta 1987, 70,
2003-2010). The reaction may be warmed to ambient temperature to
effect complete reaction. The resulting ketone is then treated with
a reducing agent, preferably sodium borohydride in a protic
solvent, preferable ethanol or methanol. Other selective reducing
reagents which will reduce the ketone but no other functional
groups, e.g., zinc borohydride or lithium triethylborohydride, can
also be employed. The temperature selection will be based upon the
activity of the reducing agent, preferably from about 0.degree. C.
to ambient temperature. The resulting hydroxyl group is then
suitably protected. Standard alcohol protecting groups such as
tetrahydropyranyl, trimethylsilyl, tert-butyl-dimethylsilyl or
benzyl may be utilized. A preferred protecting group is
tert-butyl-dimethylsilyl which is installed by standard methods as
described in Greene, T. W.; Wuts, P. G. M., Protective Groups in
Organic Synthesis, 2.sup.nd ed.; John Wiley and Sons Inc.: New
York, 1991. Preferred conditions for this reaction include treating
the alcohol in DMF at ambient temperature with 0.1 eq. of
4-dimethylaminopyridine, 1.1 eq. of
tert-butyl-dimethylsilylchloride and 2 eq. of imidazole.
The resulting compound of Formula 13 of Scheme F is then alkylated
on nitrogen with one of a variety of alkylating agents of the
formula hal-CH.sub.2CH.sub.2--X-QP to introduce the desired side
chain. The amide nitrogen is first deprotonated with a suitable
base in a reaction inert solvent. Preferred bases for this reaction
include NaN(SiMe.sub.3).sub.2 or sodium hydride in a solvent such
as DMF, tetrahydrofuran, dimethoxyethane or dioxane. An especially
preferred solvent is DMF. The appropriate temperature range for
anion formation is between -78.degree. C. and about the temperature
at which the solvent refluxes. It is preferred that the reaction is
conducted at ambient temperature. After formation of the anion, the
alkylating agent of the formula hal-CH.sub.2CH.sub.2--X-QP is
added, and the solution is stirred at a temperature between
-20.degree. C. to about the temperature at which the solvent
refluxes. A preferred temperature is between ambient temperature
and 100.degree. C. Typical alkylating agents include primary
halides and primary sulfonates. Preferably, an alkyl bromide or
alkyl iodide is used. The alcohol protecting group is then removed
by methods well known to those skilled in the art (see Greene, T.
W.; Wuts, P. G. M., Protective Groups in Organic Synthesis,
2.sup.nd ed.; John Wiley and Sons Inc.: New York, 1991) to produce
compounds of Formula 9b.
##STR00023##
Compounds of Formula 9b of Scheme F are converted to compounds of
Formula I by methods well known to those skilled in the art. In
cases where the QP group is a carboxylic ester, either acidic or
basic aqueous hydrolysis conditions may be utilized. Typically,
lower alkyl esters are hydrolyzed by base catalyzed hydrolysis in a
reaction inert solvent at ambient temperature to about the reflux
temperature of the solvent being used. Preferably the lower alkyl
ester is hydrolyzed with aqueous 1 N sodium hydroxide in methanol
at a suitable temperature, preferably at ambient temperature. When
QP is a benzyl ester or a t-butyl ester, standard deprotection
methods are utilized as described in Greene, T. W.; Wuts, P. G. M.,
Protective Groups in Organic Synthesis, 2.sup.nd ed.; John Wiley
and Sons Inc.: New York, 1991. When QP is a nitrile and not a
protected carboxylic acid, a preferred method for preparation of
the tetrazole is treatment of the nitrile with dibutyltin oxide and
trimethylsilylazide in refluxing toluene (S. J. Wittenberger and B.
G. Donner, J. Org. Chem. 1993, 58, 4139-4141). For a review of
alternative preparations of tetrazoles see R. N. Butler,
Tetrazoles, in Comprehensive Heterocyclic Chemistry; Potts, K. T.
Ed.; Pergamon Press: Oxford, 1984, Vol. 5, p 791-838.
The methods of the present invention for the treatment of
hypertension, liver failure, loss of patency of the ductus
arteriosis, glaucoma or ocular hypertension in a patient, is
demonstrated by the activity of those agonists in conventional
assays, including the prostaglandin E.sub.2 receptor subtype
binding assay, the cyclic AMP assay, and in vivo assays which
demonstrate the Formula I compounds hypotensive effect. The methods
of the present invention for the treatment of liver failure can be
demonstrated in an in vivo liver failure model. Such assays also
provide a means by which the activities of the EP.sub.4 receptor
selective agonists of Formula I can be compared with each other and
with the activity of other known compounds and compositions. The
results of these comparisons are useful for determining dosage
levels of the EP.sub.4 selective agonists of Formula I in mammals,
including humans, for the treatment of such diseases.
Administration of the selective EP.sub.4 receptor agonists
according to the methods of this invention can be via any mode that
delivers the EP.sub.4 receptor selective agonist systemically
and/or locally (e.g., at the ductus arteriosus, liver, vasculature,
or eye). These methods include oral routes, parenteral,
intraduodenal routes, etc. Generally, the compounds of this
invention are administered orally, but parenteral administration
(e.g., intravenous, intramuscular, transdermal, subcutaneous,
rectal or intramedullary) may be utilized, for example, where oral
administration is inappropriate for the target or where the patient
is unable to ingest the drug.
The methods of this invention are used for the treatment of
hypertension, liver failure, loss of patency of the ductus
arteriosus, glaucoma or ocular hypertension can be carried out by
either systemic or local application (e.g., to the ductus
arteriosus, liver, vasculature, or eye) of the selective EP.sub.4
receptor agonists. The selective EP.sub.4 receptor agonists useful
in the methods of this invention are applied to the sites of the
vasculature, liver, ductus arteriosus, or eye for example, either
by injection of the compound in a suitable solvent, or in cases of
open surgery, by local application thereto of the compound in a
suitable vehicle, carrier or diluent. In certain instances it may
be desirable to administer the selective EP.sub.4 receptor agonist
via a catheter to the site to be treated. For administration to the
eye an ophthalmic preparation, such as a gel, ointment or
ophthalmic solution or suspension can be employed.
In any event, the amount and timing of compounds administered will
be dependent on the subject being treated, on the severity of the
affliction, on the manner of administration and on the judgment of
the prescribing physician. Thus, because of patient to patient
variability, the dosages given herein are a guideline and the
physician may titrate doses of the compound to achieve the
treatment (e.g., reduce hypertension) that the physician considers
appropriate for the patient. In considering the degree of treatment
desired, the physician must balance a variety of factors such as
the age of the patient, body weight of the patient, symptom,
presence of preexisting disease, the desired therapeutic effect,
the route of administration, and the duration of the treatment,
etc. In the human adult, the dose administered is generally 1 .mu.g
to 100 mg, by oral administration, from once up to several times
per day, and from 0.1 .mu.g to 10 mg, by parenteral administration
(preferably intravenously) from once up to several times per day,
or by continuous administration for from 1 to 24 hours per day by
intravenous infusion. For the treatment of neonates the dosage will
have to be adjusted accordingly due to the patient's young age and
low body weight. In general, in the methods of the present
invention, an amount of a compound of Formula I is used that is
sufficient to treat hypertension, liver failure, loss of patency of
the ductus arteriosus, glaucoma or ocular hypertension. As the
doses to be administered depend upon various conditions, there are
cases in which doses lower or higher than the ranges specified
above can be used.
The compounds used in the methods of this invention are generally
administered in the form of a pharmaceutical composition comprising
at least one of the compounds of this invention together with a
pharmaceutically acceptable carrier, vehicle or diluent. Thus, the
selective EP.sub.4 receptor agonist can be administered
individually in any conventional local, oral, intranasal,
parenteral, rectal, topical (including ophthalmic) or transdermal
dosage form.
For oral administration the pharmaceutical composition can take the
form of solutions, suspensions, tablets, pills, capsules, powders,
and the like. Tablets containing various excipients such as sodium
citrate, calcium carbonate and calcium phosphate are employed along
with various disintegrants such as starch and preferably potato or
tapioca starch and certain complex silicates, together with binding
agents such as polyvinylpyrrolidone, sucrose, gelatin and acacia.
Additionally, lubricating agents such as magnesium stearate, sodium
lauryl sulfate and talc are often very useful for tabletting
purposes. Solid compositions of a similar type are also employed as
fillers in soft and hard-filled gelatin capsules; preferred
materials in this connection also include lactose or milk sugar as
well as high molecular weight polyethylene glycols. When aqueous
suspensions and/or elixirs are desired for oral administration, the
compositions of this invention can be combined with various
sweetening agents, flavoring agents, coloring agents, emulsifying
agents and/or suspending agents, as well as such diluents as water,
ethanol, propylene glycol, glycerin or various like combinations
thereof.
For purposes of parenteral administration, solutions in sesame or
peanut oil or in aqueous propylene glycol can be employed, as well
as sterile aqueous solutions of the corresponding water-soluble
salts. Such aqueous solutions may be suitably buffered, if
necessary, and the liquid diluent first rendered isotonic with
sufficient saline or glucose. These aqueous solutions are
especially suitable for intravenous, intramuscular, subcutaneous
and intraperitoneal injection purposes. In this connection, the
sterile aqueous media employed are all readily obtainable by
standard techniques well known to those skilled in the art.
For purposes of transdermal (e.g., topical) administration, dilute
sterile, aqueous or partially aqueous solutions (usually in about
0.1% to 5% concentration), otherwise similar to the above
parenteral solutions, are prepared.
For purposes of ophthalmic administration, an aqueous solution of
the compound of Formula I is generally preferred (typical
concentration range is 0.001 to approximately 1% weight/volume).
The aqueous solution can then be administered by instilling drops
of the solution to the patient's eyes (usually 1 to 2 drops
administered 1 to 4 times a day). For compounds of Formula I with
less water solubility, an aqueous suspension may be preferred.
Other ophthalmic compositions known in the art, such as viscous or
semi-viscous gels, or other types of solid or semi-solid
compositions containing compounds of Formula I may be employed. The
ophthalmic composition may also contain a preservative such as
benzalkonium chloride, chlorobutanol, edetate disodium, phenylethyl
alcohol, phenylmercuric acetate, phenyl mercuric nitrate, methyl
paraben, propyl paraben, polyquaternium-1, sorbic acid, thimerosal,
or other known preservatives (typical concentration range of the
preservative is 0.001 to 1.0% weight/volume). A surfactant, such as
Tween 80, can also be used in the ophthalmic composition. Various
vehicles, such as polyvinyl alcohol, povidone, hydroxypropyl methyl
cellulose, poloxamers, carboxymethyl cellulose, hydroxyethyl
cellulose cyclodextrin and water can be used for the ophthalmic
composition. The tonicity of the ophthalmic composition can be
adjusted using a tonicity adjustor such as sodium chloride,
potassium chloride, mannitol or glycerin. The ophthalmic
composition can be buffered, preferably to a range of 4.5 to 8.0,
using buffers such as acetate buffers, citrate buffers, phosphate
buffers and borate buffers. The pH of the ophthalmic composition
can be adjusted, preferably to a range between 4.5 to 8.0, using an
appropriate acid or base. Antioxidants, such as sodium
metabisulfite, sodium thiosulfate, acetylcysteine, butylated
hydroxyanisole and butylated hydroxytoluene can also be used in the
ophthalmic composition.
Methods of preparing various pharmaceutical compositions with a
certain amount of active ingredient are known, or will be apparent
in light of this disclosure, to those skilled in the art. For
examples of methods of preparing pharmaceutical compositions, see
Remington: The Science and Practice of Pharmacy, Alfonso R.
Gennaro, Mack Publishing Company, Easton, Pa., 19th Edition
(1995).
Advantageously, the present invention also provides kits for use by
a consumer to treat hypertension, liver failure, loss of patency of
the ductus arteriosus, glaucoma or ocular hypertension. The kits
comprise a) a pharmaceutical composition comprising a selective
EP.sub.4 receptor agonist of Formula I; and b) instructions
describing methods of using the pharmaceutical compositions to
treat hypertension, liver failure, loss of patency of the ductus
arteriosus, glaucoma or ocular hypertension.
A "kit" as used in the instant application includes a container for
containing the pharmaceutical compositions and may also include
divided containers such as a divided bottle or a divided foil
packet. The container can be in any conventional shape or form as
known in the art which is made of a pharmaceutically acceptable
material, for example a paper or cardboard box, a glass or plastic
bottle or jar, a re-sealable bag (for example, to hold a "refill"
of tablets for placement into a different container), or a blister
pack with individual doses for pressing out of the pack according
to a therapeutic schedule. The container employed can depend on the
exact dosage form involved, for example a conventional cardboard
box would not generally be used to hold a liquid suspension. It is
feasible that more than one container can be used together in a
single package to market a single dosage form. For example, tablets
may be contained in a bottle, which is in turn contained within a
box.
An example of such a kit is a so-called blister pack. Blister packs
are well known in the packaging industry and are being widely used
for the packaging of pharmaceutical unit dosage forms (tablets,
capsules, and the like). Blister packs generally consist of a sheet
of relatively stiff material covered with a foil of a preferably
transparent plastic material. During the packaging process,
recesses are formed in the plastic foil. The recesses have the size
and shape of individual tablets or capsules to be packed or may
have the size and shape to accommodate multiple tablets and/or
capsules to be packed. Next, the tablets or capsules are placed in
the recesses accordingly and the sheet of relatively stiff material
is sealed against the plastic foil at the face of the foil which is
opposite from the direction in which the recesses were formed. As a
result, the tablets or capsules are individually sealed or
collectively sealed, as desired, in the recesses between the
plastic foil and the sheet. Preferably, the strength of the sheet
is such that the tablets or capsules can be removed from the
blister pack by manually applying pressure on the recesses whereby
an opening is formed in the sheet at the place of the recess. The
tablet or capsule can then be removed via said opening.
It may be desirable to provide a written memory aid, where the
written memory aid is of the type containing information and/or
instructions for the physician, pharmacist or other health care
provider, or patient, e.g., in the form of numbers next to the
tablets or capsules whereby the numbers correspond with the days of
the regimen which the tablets or capsules so specified should be
ingested or a card which contains the same type of information.
Another example of such a memory aid is a calendar printed on the
card e.g., as follows "First Week, Monday, Tuesday," . . . etc. . .
. "Second Week, Monday, Tuesday, . . . " etc. Other variations of
memory aids will be readily apparent. A "daily dose" can be a
single tablet or capsule or several tablets or capsules to be taken
on a given day.
Another specific embodiment of a kit is a dispenser designed to
dispense the daily doses one at a time. Preferably, the dispenser
is equipped with a memory-aid, so as to further facilitate
compliance with the regimen. An example of such a memory-aid is a
mechanical counter that indicates the number of daily doses that
have been dispensed. Another example of such a memory-aid is a
battery-powered micro-chip memory coupled with a liquid crystal
readout, or audible reminder signal which, for example, reads out
the date that the last daily dose has been taken and/or reminds one
when the next dose is to be taken.
The kits of the present invention may also include, in addition to
a selective EP.sub.4 receptor agonist of Formula I, one or more
additional pharmaceutically active compounds. Preferably, the
additional compound is a HMG-CoA reductase inhibitor or
antihypertensive agent. The additional compound or compounds may be
administered in the same dosage form as the selective EP.sub.4
receptor agonist of Formula I or in different dosage forms.
Likewise, the additional compounds can be administered at the same
time as the selective EP.sub.4 receptor agonist of Formula I or at
different times.
In the methods of the present invention it is to be understood that
the selective EP.sub.4 receptor agonists of Formula I can be
administered in combination with other pharmaceutical agents. For
example, in the methods for treating hypertension, the selective
EP.sub.4 receptor agonists of Formula I can be administered in
combination with another antihypertensive agent. Certain patients
suffering from hypertension also suffer from other disorders such
as hypercholesterolemia or hypertriglyceridemia. In cases such as
these it is to be understood that the selective EP.sub.4 receptor
agonists of Formula I can be administered in combination with an
HMG-CoA reductase inhibitor. For patients suffering from glaucoma,
the selective EP.sub.4 receptor agonists can be administered in
combination with another anti-glaucoma agent.
Any HMG-CoA reductase inhibitor may be employed as an additional
compound in the combination therapy aspect of the present
invention. The term "HMG-CoA reductase inhibitor" or "statin"
refers to a compound that inhibits the biosynthesis of
hydroxymethylglutaryl-coenzyme A to mevalonic acid as catalyzed by
the enzyme HMG-CoA reductase. Such inhibition may be determined
readily by one of skill in the art according to standard assays
(e.g., Methods of Enzymology 1981, 71, 455-509; and the references
cited therein). A variety of these compounds are described and
referenced below. HMG-CoA reductase inhibitors may be readily
prepared by processes known in the chemical arts. Mevastatin,
lovastatin, pravastatin, velostatin, simvastatin, fluvastatin,
cerivastatin and mevastatin, dalvastatin, fluindostatin and
rivastatin may be made in accordance with the process set forth in
U.S. Pat. No. 3,983,140, U.S. Pat. No. 4,231,938, U.S. Pat. No.
4,346,227, U.S. Pat. No. 4,448,784, U.S. Pat. No. 4,450,171, U.S.
Pat. No. 4,739,073, U.S. Pat. No. 5,177,080, U.S. Pat. No.
5,177,080, European Patent Application No. 738,510 A2, European
Patent Application No. 363,934 A1 and EP 491,226 respectively.
Atorvastatin may readily be prepared as described in U.S. Pat. No.
4,681,893. The hemicalcium salt of atorvastatin, which is currently
sold as Lipitor.RTM., may readily be prepared as described in U.S.
Pat. No. 5,273,995. Other pharmaceutically-acceptable cationic
salts of atorvastatin may be readily prepared by reacting the free
acid form of atorvastatin with an appropriate base, usually one
equivalent, in a co-solvent. Other HMG-CoA reductase inhibitors
will be known to those skilled in the art. Examples of marketed
products containing HMG-CoA reductase inhibitors that can be used
in combination with compounds of Formula I in the methods of the
present invention include Lescol.RTM., Lipitor.RTM., Mevacor.RTM.,
Pravachol.RTM. and Zocor.RTM..
It is preferred that said statin is mevastatin, lovastatin,
pravastatin, velostatin, simvastatin, fluvastatin, cerivastatin,
mevastatin, dalvastatin, fluindostatin or atorvastatin, or a
prodrug thereof, or a pharmaceutically acceptable salt of said
compound or prodrug.
It is especially preferred that said statin is atorvastatin, most
preferably atorvastatin calcium.
The selective EP.sub.4 receptor agonists of Formula I can also be
administered in combination with antihypertensives in the methods
of the present invention for the treatment of hypertension.
Examples of classes of compounds that can be used to treat
hypertension (antihypertensives) include calcium channel blockers,
angiotensin converting enzyme (ACE) inhibitors, diuretics,
angiotensin II receptor blockers, .beta.-adrenergic blockers, and
.alpha.-adrenergic blockers. In addition, combinations of compounds
in the above-recited classes have been used to treat
hypertension.
Some examples of specific calcium channel blockers that can be used
in combination with the selective EP.sub.4 receptor agonists of
Formula I include amlodipine, including the besylate salt;
nifedipine; lercanidipine, verapamil, and diltiazem. Some examples
of specific .alpha.-adrenergic blockers and related compounds
include doxazosin, including the mesylate salt; prazosin, including
the hydrochloride salt; and prazosin hydrochloride/polythiazide.
Some examples of specific .beta.-adrenergic blockers that can be
used in combination with the selective EP.sub.4 receptor agonists
of Formula I include sotalol, including the hydrochloride salt;
timolol, including the maleate salt; propanolol, including the
hydrochloride salt; acebutolol, including its hydrochloride salt;
betaxolol, including the hydrochloride salt; penbutolol, including
its sulfate salt; nadolol; bisoprolol, including the fumarate salt;
atenolol; and metoprolol, including the succinate salt. Angiotensin
II inhibitors such as candesartan cilexetil, irbesartan, losartan
potassium, valsartan, and telmisartan can also be used in
combination with the selective EP.sub.4 receptor agonists of
Formula I. Diuretics such as carbonic anhydrase inhibitors,
combination diuretics, loop diuretics, potassium-sparing diuretics
and thiazide and related diuretics can be used in combination with
the compounds of Formula I. Some examples of specific diuretics
that can be used in combination with the compounds of Formula I
include hydrochlorothiazide, dichlorphenamide, spironolactone with
hydrochlorothiazide, triamterene, hydrochlorothiazide with
triamterene, amiloride hydrochloride, amiloride hydrochloride with
hydrochlorothiazide, torsemide, ethacrynic acid, furosemide,
hydroflumethazide, chlorothiazide, methyclothiazide, indapamide,
metolazone, polythiazide and chlorthalidone. Some examples of
specific ACE inhibitors including quinapril, captopril, alacepril,
moveltipril, zofenopril, enalapril, enalaprilat, delapril,
ramipril, spirapril, lisinopril, benazepril, cilazapril,
perindopril, fosinopril and trandolapril can also be used in
combination with the selective EP.sub.4 receptor agonists of
Formula I.
Combination therapy can also be used in the methods of the present
invention for the treatment of glaucoma or ocular hypertension. For
the treatment of glaucoma or ocular hypertension, the selective
EP.sub.4 receptor agonists of Formula I can be combined with other
medicaments known to be useful for the treatment of glaucoma
(anti-glaucoma agents), such as .beta.-adrenergic blocking agents,
carbonic anhydrase inhibitors, miotics and sympathomimetics. For
example, .beta.-adrenergic agents such as betaxolol, including its
hydrochloride salt, and timolol, including its maleate salt can be
combined with the selective EP.sub.4 receptor agonists of Formula
I. Some examples of specific carbonic anhydrase inhibitors that can
be used in combination with the selective EP.sub.4 receptor
agonists of Formula I include brinzolamide, dichlorphenamide, and
dorzolamide, including its hydrochloride salt. Miotics, such as
demecarium bromide, can also be used in combination with the
selective EP.sub.4 receptor agonists of Formula I.
Sympathomimetics, such as brimonidine, including its tartrate salt,
pheniramine, including its maleate salt, and phenylephrine,
including its hydrochloride salt, can be used in combination with
the selective EP.sub.4 receptor agonists of Formula I.
In the combination therapy aspect of the methods of the present
invention, the selective EP.sub.4 receptor agonists of Formula I
and any additional compounds, such as the HMG-CoA reductase
inhibitors and/or antihypertensive agents or the anti-glaucoma
agents, can be administered in the same dosage form or in separate
dosage forms. The dosage forms can be the same (e.g., both tablets)
or different. Likewise, the compounds can be administered at the
same time or at different times. All variations are intended to be
included in the methods of the present invention.
The documents cited herein, including any patents and patent
applications, are hereby incorporated by reference.
EXPERIMENTAL SECTION
General Experimental Procedures
NMR spectra were recorded on a Varian Unity 400 spectrometer
(Varian Co., Palo Alto, Calif.) at about 23.degree. C. at 400 MHz
for proton nuclei. Chemical shifts are expressed in parts per
million. The peak shapes are denoted as follows: s, singlet; d,
doublet; t, triplet; q, quartet; m, multiplet; bs, broad singlet.
Atmospheric pressure chemical ionization (APCI) mass spectra were
obtained on a Fisons Platform II Spectrometer (Micromass Inc.,
Beverly, Mass.). Where the intensity of chlorine or
bromine-containing ions are described the expected intensity ratio
was observed (approximately 3:1 for .sup.35Cl/.sup.37Cl-containing
ions) and 1:1 for .sup.79Br/.sup.81Br-containing ions) and the
intensity of only the lower mass ion is given.
Medium pressure chromatography was performed using a Biotage
purification system (Biotage, Dyax Corporation, Charlottesville,
Va.) under nitrogen pressure. Flash chromatography was performed
with either Baker Silica Gel (40 .mu.m) (J. T. Baker, Phillipsburg,
N.J.) or Silica Gel 60 (EM Sciences, Gibbstown, N.J.) in glass
columns under low nitrogen pressure. Radial Chromatography was
performed using a Chromatotron (Harrison Research, Palo Alto,
Calif.). Preparative Chromatography was performed using Analtech
Uniplates Silica Gel GF (20.times.20 cm) (Analtech, Inc. Newark,
Del.). Dimethylformamide (DMF), tetrahydrofuran (THF), and
dichloromethane (CH.sub.2Cl.sub.2) used as reaction solvents were
the anhydrous grade supplied by Aldrich Chemical Company
(Milwaukee, Wis.). The term "concentrated" refers to removal of
solvent at water aspirator pressure on a rotary evaporator. The
term "EtOAc" means ethyl acetate. The term "Et.sub.2O" means
diethyl ether. The term "MeOH" means methanol. The abbreviation `h`
stands for hours. The term "TBAF" refers to tetrabutylammonium
fluoride. The term "DMAP" refers to dimethylaminopyridine. The
terms "dichloromethane" and "methylene chloride" are synonymous and
are used interchangeably throughout this description and in the
following compounds and preparations section. The following section
describes preparations and compounds of Formula I. The compounds
described below can be used in the methods of the present
invention.
The examples presented herein are intended to illustrate particular
embodiments of the invention, and are not intended to limit the
specification or the claims in any manner.
Preparation of Specific Embodiments of Formula I Compounds
The following section provides specific embodiments of Formula I
compounds (Compounds 1A-1H, 2A-2K, 3A-3M, 4A-4B, and 5A-5B) and
preparations (Preparations 1-26) useful for their synthesis. The
compounds provided can be used in the methods of the present
invention.
Compound 1A
4-{3-[2-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-benzoic
acid
Step A: 5-(3-Oxo-4-phenyl-butyl)-pyrrolidin-2-one. To a solution of
tetrahydro-pyrrolizine-3,5-dione (5 g, 36 mmol) in CH.sub.2Cl.sub.2
(320 mL) at 0.degree. C. was added benzyl magnesium chloride (1M
solution in THF, 39 mL, 39 mmol) dropwise. The solution was stirred
at 0.degree. C. for 3 h and was quenched with saturated aqueous
ammonium chloride. After warming to room temperature, the aqueous
solution was extracted with CH.sub.2Cl.sub.2 (3.times.). The
combined organic extracts were dried (MgSO.sub.4), filtered and
concentrated. The residue was purified by medium pressure
chromatography eluting with a solvent gradient (1% MeOH in
CH.sub.2Cl.sub.2 to 2% MeOH in CH.sub.2Cl.sub.2) to yield 5.9021 g
of 5-(3-oxo-4-phenyl-butyl)-pyrrolidin-2-one. .sup.1H NMR
(CDCl.sub.3) .delta. 87.35-7.18 (m, 5H), 3.69 (s, 2H), 3.56 (m,
1H), 2.50 (t, 2H), 2.27 (m, 2H), 2.15 (m, 1H), 1.73 (m, 2H), 1.61
(m, 1H).
Step B: 5-(3-Hydroxy-4-phenyl-butyl)-pyrrolidin-2-one. To a
solution of 5-(3-oxo-4-phenyl-butyl)-pyrrolidin-2-one (5.902 g,
25.52 mmol) in EtOH (30 mL) at 0.degree. C. was added NaBH.sub.4
(485 mg, 12.76 mmol) and the reaction mixture was stirred at
0.degree. C. for 2.5 h. The reaction mixture was quenched with
saturated aqueous ammonium chloride. Water and CH.sub.2Cl.sub.2
were added. The aqueous layer was washed with CH.sub.2Cl.sub.2
(2.times.) and the combined organic extracts were dried
(MgSO.sub.4), filtered and concentrated. The residue was purified
by medium pressure chromatography with a solvent gradient (1:1
hexanes:EtOAc to EtOAc to 1% MeOH in CH.sub.2Cl.sub.2) to yield 4.3
g of 5-(3-hydroxy-4-phenyl-butyl)-pyrrolidin-2-one. .sup.1H NMR
(CDCl.sub.3) .delta. 7.35-7.16 (m, 5H), 6.02 (m, 1H), 3.80 (m, 1H),
3.63 (m, 1H), 2.79 (m, 1H), 2.64 (m, 1H), 2.26 (m, 3H), 1.72-1.22
(m, 6H).
Step C:
5-[3-(tert-Butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-pyrrolidin--
2-one. To a solution of
5-(3-hydroxy-4-phenyl-butyl)-pyrrolidin-2-one (4.3 g, 18.43 mmol)
in DMF (86 mL) was added tert-butyldimethylsilyl chloride (3.06 g,
20.3 mmol) followed by imidazole (2.5 g, 37 mmol) and DMAP (225
mg). The reaction mixture was stirred for 24 h and was quenched
with saturated aqueous ammonium chloride. The aqueous solution was
washed with EtOAc (3.times.) and the combined organic extracts were
dried (MgSO.sub.4), filtered and concentrated. The residue was
purified by medium pressure chromatography eluting with a solvent
gradient (CH.sub.2Cl.sub.2 to 1% MeOH in CH.sub.2Cl.sub.2 to 2%
MeOH in CH.sub.2Cl.sub.2) to yield 5.94 g of
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-pyrrolidin-2-one.
.sup.1H NMR (CDCl.sub.3) .delta. 7.26-7.10 (m, 5H), 5.68 (m, 1H),
3.83 (m, 1H), 3.54 (m, 1H), 2.69 (m, 2H), 2.30-2.16 (m, 3H),
1.66-1.35 (m, 5H), 0.82 (s, 9H), -0.06 (d, 3H), -0.2 (d, 3H).
Step D:
4-(3-{2-[3-(tert-Butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-5-oxo-
-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester. To a solution
of
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-pyrrolidin-2-one
(3.20 g, 9.21 mmol) in DMF (30 mL) at 0.degree. C. was added NaHMDS
(1M in THF, 11.5 mL, 11.5 mmol). After 1 h,
4-(3-bromo-propyl)-benzoic acid methyl ester (2.84 g, 11.0 mmol)
was added and the reaction mixture was stirred at 70.degree. C. for
18 h. The DMF was removed in vacuo and the residue was dissolved in
EtOAc. The organic solution was washed with water, dried
(MgSO.sub.4), filtered and concentrated. The residue was purified
by medium pressure chromatography (30% EtOAc in hexanes) to yield
3.39 g of
4-(3-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-5-oxo-pyrroli-
din-1-yl}-propyl)-benzoic acid methyl ester. .sup.1H NMR
(CDCl.sub.3) (selected peaks) .delta. 7.92 (m, 2H), 7.25-7.09 (m,
7H), 3.86 (s, 3H), 3.80 (m, 1H), 3.61 (m, 1H), 3.46 (m, 1H), 2.90
(m, 1H), 2.78-2.57 (m, 4H), 2.38-2.18 (m, 2H), 0.83 (s, 9H); MS
524.1 (M+1).
Step E:
4-{3-[2-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-
-benzoic acid methyl ester. To a solution of
4-(3-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-5-oxo-pyrroli-
din-1-yl}-propyl)-benzoic acid methyl ester (3.37 g, 6.43 mmol) in
THF (40 mL) at 0.degree. C. was added tetra-butylammonium fluoride
(1M in THF, 9.6 mL, 9.6 mmol). The reaction mixture was stirred at
room temperature for 18 h and the volatiles were removed in vacuo.
EtOAc was added and the organic solution was washed with saturated
aqueous NaHCO.sub.3 (2.times.), water (1.times.), and brine
(1.times.). The organic solution was dried (MgSO.sub.4), filtered
and concentrated. The residue was purified by medium pressure
chromatography eluting with EtOAc to yield 2.28 g of
4-{3-[2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propy-
l}-benzoic acid methyl ester. .sup.1H NMR (CDCl.sub.3) (selected
peaks) .delta. 7.91 (d, 2H), 7.32-7.15 (m, 7H), 3.86 (s, 3H), 3.75
(m, 1H), 3.63 (m, 1H), 3.54 (m, 1H), 2.94 (m, 1H), 2.78 (m, 1H),
2.61 (m, 3H); MS 410.1 (M+1).
Step F:
4-{3-[2-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-
-benzoic acid. To a solution of
4-{3-[2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-benzoic
acid methyl ester (2.28 g, 5.57 mmol) in MeOH (20 mL) was added 2N
NaOH (5 mL). The reaction mixture was stirred at room temperature
for 20 h and was heated under reflux for 3 h. The volatiles were
removed in vacuo and the residue was diluted with CH.sub.2Cl.sub.2
and 1N HCl. The aqueous solution was extracted with
CH.sub.2Cl.sub.2 (2.times.) and the combined organic extracts were
washed with brine. The organic solution was dried (MgSO.sub.4),
filtered and concentrated to yield the title compound (2.03 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.98 (d, 2H), 7.34-7.18 (m, 7H),
3.80 (m, 1H), 3.67 (m, 1H), 3.58 (m, 1H), 2.97 (m, 1H), 2.81 (m,
1H), 2.68 (m, 3H), 2.45-2.27 (m, 2H), 2.13-1.30 (m, 9H); MS 396.3
(M+1), 394.2 (M-1).
Compound 1B
4-(3-{2-[3-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-1--
yl}-propyl)-benzoic acid
Step A:
5-[3-Oxo-4-(3-trifluoromethyl-phenyl)-butyl]-pyrrolidin-2-one
Magnesium coils (1.13 g) were stirred under vacuum in a round
bottom flask for 60 h. Anhydrous Et.sub.2O (5 mL) was added and the
reaction mixture was cooled to 0.degree. C. A solution of
3-trifluoromethylbenzyl chloride (1.0 mL, 7.5 mmol) in Et.sub.2O
(25 mL) was added dropwise over 3 h. The reaction mixture was
stirred for an additional 2.5 h. The solution was slowly added via
a syringe and filtered through a nylon syringe filter into a
solution of tetrahydro-pyrrolizine-3,5-dione (650 mg, 4.68 mmol) in
CH.sub.2Cl.sub.2 (30 mL) at 0.degree. C. After 2 h, the reaction
mixture was quenched with 1N HCl and the aqueous solution was
washed with CH.sub.2Cl.sub.2 (2.times.). The organic solutions were
combined, dried (MgSO.sub.4), filtered and concentrated. Medium
pressure chromatography (1:1 hexanes:EtOAc) provided
5-[3-oxo-4-(3-trifluoromethyl-phenyl)-butyl]-pyrrolidin-2-one
(1.376 g). .sup.1H NMR (CDCl.sub.3) .delta. 7.38 (m, 4H), 3.78 (s,
2H), 3.61 (m, 1H), 2.58 (t, 2H), 2.30 (m, 2H), 2.20 (m, 1H),
2.86-1.59 (m, 3H).
Step B:
5-[3-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-pyrrolidin-2-one-
. Analogous to the procedure described for Compound 1A, Step B,
5-[3-oxo-4-(3-trifluoromethyl-phenyl)-butyl]-pyrrolidin-2-one (1.37
g, 4.59 mmol) was reduced with NaBH.sub.4 (174 mg) at 0.degree. C.
over 2 h. Purification by medium pressure chromatography (2% MeOH
in CH.sub.2Cl.sub.2) provided
5-[3-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-pyrrolidin-2-one
(1.19 g). .sup.1H NMR (CDCl.sub.3) .delta. 7.42 (m, 4H), 6.26 (m,
1H), 3.82 (m, 1H), 3.65 (m, 1H), 2.84 (m, 1H), 2.72 (m, 1H), 2.27
(m, 3H), 1.86 (m, 1H), 1.75-1.42 (m, 5H); MS 302.2 (M+1).
Step C:
5-[3-(tert-Butyl-dimethyl-silanyloxy)-4-(3-trifluoromethyl-phenyl-
)-butyl]-pyrrolidin-2-one. Analogous to the procedure described for
Compound 1A, Step C,
5-[3-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-pyrrolidin-2-one
(1.19 g, 3.95 mmol) was protected with tert-butyidimethylsilyl
chloride (893 mg, 6.22 mmol). Purification by medium pressure
chromatography eluting with EtOAc provided
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-trifluoromethyl-phenyl)-butyl]-
-pyrrolidin-2-one. .sup.1H NMR (CDCl.sub.3) .delta.7.47-7.32 (m,
4H), 5.73 (m, 1H), 3.86 (m, 1H), 3.59 (m, 1H), 2.75 (m, 2H),
2.35-2.20 (m, 3H), 1.70-1.40 (m, 5H), 0.81 (s, 9H), -0.05 (d, 3H),
-0.3 (d, 3H); MS 416.1 (M+1).
Step D:
4-(3-{2-[3-(tert-Butyl-dimethyl-silanyloxy)-4-(3-trifluoromethyl--
phenyl)-butyl]-5-oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl
ester. Analogous to the procedure described for Compound 1A, Step
D,
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-trifluoromethyl-phenyl)-butyl]-
-pyrrolidin-2-one (250 mg, 0.602 mmol) was alkylated with NaHMDS
(1M in THF, 0.72 mL, 0.72 mmol) and 4-(3-bromo-propyl)-benzoic acid
methyl ester (170 mg, 0.663 mmol) to yield
4-(3-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-trifluoromethyl-phenyl)--
butyl]-5-oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester
(300 mg). MS 592.1 (M+1).
Step E:
4-(3-{2-[3-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrr-
olidin-1-yl}-propyl)-benzoic acid methyl ester.
4-(3-{2-[3-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-benzoic acid methyl ester was prepared Analogous to
the procedure described for Compound 1A, Step E. .sup.1H NMR
(CDCl.sub.3) (selected peaks) .delta. 7.91 (d, 2H), 7.49-7.35 (m,
4H), 7.22 (d, 2H), 3.85 (s, 3H), 3.80 (m, 1H), 3.65 (m, 1H), 3.55
(m, 1H), 2.98-2.61 (m, 5H).
Step F:
4-(3-{2-[3-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrr-
olidin-1-yl}-propyl)-benzoic acid. Analogous to the procedure
described for Compound 1A, Step F,
4-(3-{2-[3-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-1-
-yl}propyl)-benzoic acid methyl ester was hydrolyzed at room
temperature over 24 h to generate
4-(3-{2-[3-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-benzoic acid. .sup.1H NMR (CDCl.sub.3) .delta. 7.98
(d, 2H), 7.52-7.37 (m, 4H), 7.26 (d, 2H), 3.82 (m, 1H), 3.68 (m,
1H), 3.58 (m, 1H), 2.98-2.66 (m, 5H), 2.34 (m, 2H), 2.09 (m, 1H),
1.95-1.37 (m, 7H); MS 464.2 (M+1).
Compound 1C
4-(3-{2-[4-(3-Chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-propy-
l)-benzoic acid
Step A: 5-[4-(3-Chloro-phenyl)-3-oxo-butyl]-pyrrolidin-2-one.
Analogous to the procedure described for Compound 1A, Step A,
tetrahydro-pyrrolizine-3,5-dione (2 g, 14 mmol) was reacted with
3-chlorobenzylmagnesium chloride (0.25M in Et.sub.2O, 62 mL, 15.5
mmol) over 2 h. Purification by medium pressure chromatography
eluting with a solvent gradient (2:1 hexanes:EtOAc to EtOAc to 5%
MeOH in CH.sub.2Cl.sub.2) provided
5-[4-(3-chloro-phenyl)-3-oxo-butyl]-pyrrolidin-2-one (1.9142 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.27 (m, 2H), 7.19 (m, 1H), 7.08
(m, 1H), 6.27 (br, 1H), 3.68 (s, 2H), 3.60 (m, 1H), 2.52 (t, 2H),
2.29 (m, 2H), 2.21 (m, 1H), 1.88-1.60 (m, 3H); MS 266.2 (M+1),
264.2 (M-1).
Step B: 5-[4-(3-Chloro-phenyl)-3-hydroxy-butyl]-pyrrolidin-2-one.
Analogous to the procedure described for Compound 1A, Step B,
5-[4-(3-chloro-phenyl)-3-oxo-butyl]-pyrrolidin-2-one (1.9 g, 7.15
mmol) was reduced with NaBH.sub.4 (135 mg, 3.57 mmol). Purification
by medium pressure chromatography eluting with a solvent gradient
(1:1 hexanes:EtOAc to EtOAc to 1% MeOH in CH.sub.2Cl.sub.2 to 4%
MeOH in CH.sub.2Cl.sub.2 to 8% MeOH in CH.sub.2Cl.sub.2) provided
5-[4-(3-chloro-phenyl)-3-hydroxy-butyl]-pyrrolidin-2-one (1.53 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.22 (m, 3H), 7.07 (m, 1H), 6.51
(d, 1H), 3.82 (m, 1H), 3.66 (m, 1H), 2.77 (m, 1H), 2.66 (m, 1H),
2.33-2.19 (m, 3H), 2.04 (d, 1H), 1.74-1.45 (m, 5H); MS 268.2
(M+1).
Step C:
5-[3-(tert-Butyl-dimethyl-silanyloxy)-4-(3-chloro-phenyl)-butyl]--
pyrrolidin-2-one. Analogous to the procedure described for Compound
1A, Step C,
5-[4-(3-chloro-phenyl)-3-hydroxy-butyl]-pyrrolidin-2-one (1.53 g,
5.71 mmol) was reacted with tert-butyidimethylsilyl chloride (0.97
g, 6.4 mmol). Purification by medium pressure chromatography using
a solvent gradient (1:1 hexanes:EtOAc to EtOAc to 1% MeOH in
CH.sub.2Cl.sub.2 to 2% MeOH in CH.sub.2Cl.sub.2 to 4% MeOH in
CH.sub.2Cl.sub.2) provided
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-chloro-phenyl)-butyl]-pyrrolid-
in-2-one (1.77 g). .sup.1H NMR (CDCl.sub.3) .delta.7.16 (m, 3H),
7.01 (m, 1H), 5.61 (d, 1H), 3.83 (m, 1H), 3.58 (m, 1H), 2.68 (m,
2H), 2.28 (m, 3H), 1.73-1.36 (m, 5H), 0.84 (s, 9H), -0.05 (s, 3H),
-0.2 (d, 3H).
Step D:
4-(3-{2-[3-(tert-Butyl-dimethyl-silanyloxy)-4-(3-chloro-phenyl)-b-
utyl]-5-oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester.
Analogous to the procedure described for Compound 1A, Step D,
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-chloro-phenyl)-butyl]-pyrrolid-
in-2-one (246.5 mg, 0.645 mmol) was alkylated with NaHMDS (1M in
THF, 0.77 mL, 0.77 mmol) and 4-(3-bromo-propyl)-benzoic acid methyl
ester (200 mg, 0.767 mmol). Purification by medium pressure
chromatography (5:1 hexanes:EtOAc to 1:1 hexanes:EtOAc to EtOAc to
1% MeOH in CH.sub.2Cl.sub.2 to 5% MeOH in CH.sub.2Cl.sub.2)
provided
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-chloro-phenyl)-butyl]-pyrrolid-
in-2-one (246.3 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.94 (d, 2H),
7.25-7.13 (m, 5H), 7.01 (m, 1H), 3.88 (s, 3H), 3.82 (m, 1H), 3.66
(m, 1H), 3.50 (m, 1H), 2.94 (m, 1H), 2.73-2.57 (m, 4H), 2.47-2.27
(m, 2H), 2.12-11.23 (m, 8H), 0.84 (s, 9H), -0.05 (d, 3H), -0.2 (d,
3H); MS 558.5 (M+).
Step E:
4-(3-{2-[4-(3-Chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1--
yl}-propyl)-benzoic acid methyl ester.
4-(3-{2-[4-(3-Chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-benzoic acid methyl ester was prepared Analogous to the
procedure described for Compound 1A, Step E after purification by
medium pressure chromatography (CH.sub.2Cl.sub.2 to 1% MeOH in
CH.sub.2Cl.sub.2 to 2% MeOH in CH.sub.2Cl.sub.2 to 5% MeOH in
CH.sub.2Cl.sub.2). .sup.1H NMR (CDCl.sub.3) .delta. 7.94 (d, 2H),
7.25-7.19 (m, 5H), 7.07 (m, 1H), 3.88 (s, 3H), 3.78 (m, 1H), 3.66
(m, 1H), 3.58 (m, 1H), 2.97 (m, 1H), 2.76 (m, 1H), 2.68-2.58 (m,
3H), 2.45-2.27 (m, 2H), 2.07 (m, 1H), 1.95-1.34 (m, 8H).
Step F:
4-(3-{2-[4-(3-Chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1--
yl}-propyl)-benzoic acid. Analogous to the procedure described for
Compound 1A, Step F,
4-(3-{2-[4-(3-chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-benzoic acid methyl ester was hydrolyzed with 6N NaOH at room
temperature over 24 h to generate
4-(3-{2-[4-(3-chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-benzoic acid. .sup.1H NMR (CDCl.sub.3) .delta. 7.98 (d, 2H),
7.27-7.09 (m, 6H), 3.81 (m, 1H), 3.65 (m, 2H), 2.99 (m, 2H), 2.75
(m, 3H), 2.39 (m, 2H), 2.20-1.30 (m, 9H).
Compound 1D
4-(3-{2-[4-(3-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-propy-
l)-benzoic acid
Step A: 5-[4-(3-Fluoro-phenyl)-3-oxo-butyl]-pyrrolidin-2-one.
Analogous to the procedure described for Compound 1A, Step A,
tetrahydro-pyrrolizine-3,5-dione (2 g, 14 mmol) was reacted with
3-fluorobenzylmagnesium chloride (0.25M in Et.sub.2O, 62 mL, 15.5
mmol) over 2.5 h. Purification by medium pressure chromatography
using a solvent gradient (1:1 hexanes:EtOAc to 2:1 EtOAc:hexanes to
EtOAc to 2% MeOH in CH.sub.2Cl.sub.2 to 10% MeOH in
CH.sub.2Cl.sub.2) provided
5-[4-(3-fluoro-phenyl)-3-oxo-butyl]-pyrrolidin-2-one (2.1730 g).
.sup.1 H NMR (CDCl.sub.3) .delta. 7.32-7.27 (m, 1H), 7.00-6.90 (m,
3H), 6.12 (bs, 1H), 3.69 (s, 2H), 3.59 (m, 1H), 2.52 (t, 2H), 2.30
(m, 2H), 2.19 (m, 1H), 1.75 (m, 2H), 1.65 (m, 1H).
Step B: 5-[4-(3-Fluoro-phenyl)-3-hydroxy-butyl]-pyrrolidin-2-one.
Analogous to the procedure described for Compound 1A, Step B,
5-[4-(3-fluoro-phenyl)-3-oxo-butyl]-pyrrolidin-2-one (2.17 g, 8.71
mmol) was reduced with NaBH.sub.4 (165 mg, 4.35 mmol). Purification
by medium pressure chromatography using a solvent gradient (1:1
hexanes:EtOAc to EtOAc to 1% MeOH in CH.sub.2Cl.sub.2 to 3% MeOH in
CH.sub.2Cl.sub.2 to 6% MeOH in CH.sub.2Cl.sub.2) provided
5-[4-(3-fluoro-phenyl)-3-hydroxy-butyl]-pyrrolidin-2-one (2.23 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.27 (m, 1H), 6.94 (m, 3H), 6.38
(m, 1H), 3.82 (m, 1H), 3.66 (m, 1H), 2.79 (m, 1H), 2.67 (m, 1H),
2.33-2.21 (m, 3H), 1.92 (d, 1H), 1.75-1.40 (m, 5H); MS 252.2
(M+1).
Step C:
5-[3-(tert-Butyl-dimethyl-silanyloxy)-4-(3-fluoro-phenyl)-butyl]--
pyrrolidin-2-one. Analogous to the procedure described for Compound
1A, Step C,
5-[4-(3-fluoro-phenyl)-3-hydroxy-butyl]-pyrrolidin-2-one (2.23 g,
8.87 mmol) was reacted with tert-butyldimethylsilyl chloride (1.47
g, 9.76 mmol). Purification by medium pressure chromatography using
a solvent gradient (1:1 hexanes:EtOAc to EtOAc to 1% MeOH in
CH.sub.2Cl.sub.2 to 2% MeOH in CH.sub.2Cl.sub.2 to 4% MeOH in
CH.sub.2Cl.sub.2) provided
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-fluoro-phenyl)-butyl]-pyrrolid-
in-2-one (2.84 g). .sup.1H NMR (CDCl.sub.3) .delta. 7.23 (m, 1H),
6.88 (m, 3H), 5.75 (m, 1H), 3.85 (m, 1H), 3.57 (m, 1H), 2.71 (m,
2H), 2.30 (m, 2H), 2.25 (m, 1H), 1.70-1.38 (m, 5H), 0.84 (s, 9H), 0
(s, 3H), -0.2 (s, 3H).
Step D:
4-(3-{2-[3-(tert-Butyl-dimethyl-silanyloxy)-4-(3-fluoro-phenyl)-b-
utyl]-5-oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester.
Analogous to the procedure described in Compound 1A, Step D,
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-fluoro-phenyl)-butyl]-pyrrolid-
in-2-one (254.7 mg, 0.697 mmol) was alkylated with NaHMDS (1M in
THF, 0.84 mL, 0.84 mmol) and 4-(3-bromo-propyl)-benzoic acid methyl
ester (200 mg, 0.778 mmol). Purificaton by medium pressure
chromatography (5:1 hexanes:EtOAc to 1:1 hexanes:EtOAc to EtOAc to
1% MeOH in CH.sub.2Cl.sub.2 to 5% MeOH in CH.sub.2Cl.sub.2)
provided
4-(3-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-fluoro-phenyl)-butyl]-5--
oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester (275.3 mg).
.sup.1H NMR (CDCl.sub.3) (selected peaks) .delta. 7.94 (d, 2H),
7.23 (m, 3H), 6.87 (m, 3H), 3.88 (s, 3H), 3.86 (m, 1H), 3.63 (m,
1H), 3.50 (m, 1H), 2.94 (m, 1H), 0.84 (s, 9H).
Step E:
4-(3-{2-[4-(3-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1--
yl}-propyl)-benzoic acid methyl ester. Analogous to the procedure
described for Compound 1A, Step E,
4-(3-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-fluoro-phenyl)-butyl]-5--
oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester (275.3 mg,
0.508 mmol) was deprotected to yield
4-(3-{2-[4-(3-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-benzoic acid methyl ester (217.2 mg). Purification was
performed by medium pressure chromatography eluting with a solvent
gradient (CH.sub.2Cl.sub.2 to 1% MeOH in CH.sub.2Cl.sub.2 to 2%
MeOH in CH.sub.2Cl.sub.2 to 5% MeOH in CH.sub.2Cl.sub.2). .sup.1H
NMR (CDCl.sub.3) .delta. 7.94 (d, J=7.88 Hz, 2H), 7.27 (m, 3H),
6.93 (m, 3H), 3.88 (s, 3H), 3.78 (m, 1H), 3.66 (m, 1H), 3.57 (m,
1H), 2.97 (m, 1H), 2.78 (m, 1H), 2.64 (m, 4H), 2.45-2.25 (m, 2H),
2.07 (m, 1H), 1.95-1.30 (m, 7H).
Step F:
4-(3-{2-[4-(3-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1--
yl}-propyl)-benzoic acid. Analogous to the procedure described for
Compound 1A, Step F,
4-(3-{2-[4-(3-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-benzoic acid methyl ester was hydrolyzed with 6N NaOH at room
temperature over 24 h to generate
4-(3-{2-[4-(3-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-benzoic acid. .sup.1H NMR (CDCl.sub.3) .delta. 7.99 (d, 2H),
7.26 (m, 3H), 6.95 (m, 3H), 3.81 (m, 1H), 3.65 (m, 2H), 3.01 (m,
1H), 2.86-2.66 (m, 3H), 2.39 (m, 2H), 2.08 (m, 1H), 200-1.30 (m,
9H).
Compound 1E
4-(3-{2-[3-Hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-benzoic acid
Step A: 5-[3-Oxo-4-(3-phenoxy-phenyl)-butyl]-pyrrolidin-2-one.
Analogous to the procedure described for Compound 1B, Step A,
tetrahydro-pyrrolizine-3,5-dione (650 mg, 4.68 mmol) and
3-phenoxybenzyl chloride (1.20 g, 5.49 mmol) were reacted over 3.5
h to provide 5-[3-oxo-4-(3-phenoxy-phenyl)-butyl]-pyrrolidin-2-one
(924 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.30 (m, 3H), 7.10 (m,
1H), 6.99 (m, 2H), 6.92-6.84 (m, 3H), 3.66 (s, 2H), 3.57 (m, 1H),
2.52 (t, 2H), 2.27 (m, 2H), 2.17 (m, 1H), 1.80-1.58 (m, 3H).
Step B: 5-[3-Hydroxy-4-(3-phenoxy-phenyl)-butyl]-pyrrolidin-2-one.
Analogous to the procedure described for Compound 1A, Step B,
5-[3-oxo-4-(3-phenoxy-phenyl)-butyl]-pyrrolidin-2-one (923.6 mg,
2.86 mmol) was reduced with NaBH.sub.4 (54 mg, 1.4 mmol).
Purification by medium pressure chromatography (1:1 hexanes:EtOAc
to 2% MeOH in CH.sub.2Cl.sub.2 to 4% MeOH in CH.sub.2Cl.sub.2 to
10% MeOH in CH.sub.2Cl.sub.2) provided
5-[3-hydroxy-4-(3-phenoxy-phenyl)-butyl]-pyrrolidin-2-one (668.3
mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.31 (m, 2H), 7.23 (m, 1H),
7.08 (m, 1H), 6.97 (d, 2H), 6.91 (d, 1H), 6.84 (m, 2H), 3.80 (m,
1H), 3.73 (m, 1H), 2.77-2.03 (m, 2H), 2.40 (m, 2H), 2.24 (m, 1H),
1.75-1.41 (m, 5H); MS 326.3 (M+1).
Step C:
5-[3-(tert-Butyl-dimethyl-silanyloxy)-4-(3-phenoxy-phenyl)-butyl]-
-pyrrolidin-2-one. Analogous to the procedure described for
Compound 1A, Step C,
5-[3-hydroxy-4-(3-phenoxy-phenyl)-butyl]-pyrrolidin-2-one (668.3
mg, 2.05 mmol) was reacted with tert-butyldimethylsilyl chloride
(341 mg, 2.26 mmol). Purification by medium pressure chromatography
(CH.sub.2Cl.sub.2 to 1% MeOH in CH.sub.2Cl.sub.2 to 2% MeOH in
CH.sub.2Cl.sub.2) provided
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-phenoxy-phenyl)-butyl]-pyrroli-
din-2-one (673 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.32 (m, 2H),
7.22 (m, 1H), 7.09 (m, 1H), 6.99 (d, 2H), 6.89 (d, 1H), 6.83 (m,
2H), 3.85 (m, 1H), 3.58 (m, 1H), 2.76-2.62 (m, 2H), 2.32 (m, 2H),
2.23 (m, 1H), 1.73-1.34 (m, 5H), 0.84 (s, 9H), -0.03 (d, 3H), -0.16
(d, 3H); MS 440.7 (M+1).
Step D:
4-(3-{2-[3-(tert-Butyl-dimethyl-silanyloxy)-4-(3-phenoxy-phenyl)--
butyl]-5-oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester.
Analogous to the procedure described for Compound 1A, Step D,
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-phenoxy-phenyl)-butyl]-pyrroli-
din-2-one (200 mg, 0.455 mmol) was alkylated with NaHMDS (1M in
THF, 0.55 mL, 0.55 mmol) and 4-(3-bromo-propyl)-benzoic acid methyl
ester (128 mg, 0.501 mmol) to yield
4-(3-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-(3-phenoxy-phenyl)-butyl]-5-
-oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester (173.1 mg).
.sup.1H NMR (CDCl.sub.3) .delta. 7.94 (d, 2H), 7.32 (m, 2H),
7.25-7.19 (m, 3H), 7.09 (m, 1H), 6.98 (d, 2H), 6.88-6.81 (m, 3H),
3.88 (s, 3H), 3.84 (m, 1H), 3.64 (m, 1H), 3.50 (m, 1H), 2.95 (m,
1H), 2.76-2.57 (m, 4H), 2.37 (m, 2H), 2.03 (m, 1H), 1.92-1.67 (m,
3H), 1.56 (m, 1H), 1.46-1.25 (m, 3H), 0.84 (s, 9H), -0.04 (d, 3H),
-0.15 (d, 3H).
Step E:
4-(3-{2-[3-Hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-benzoic acid methyl ester.
4-(3-{2-[3-Hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-benzoic acid methyl ester was prepared analogous to the
procedure described for Compound 1A, Step E after purification by
medium pressure chromatography (CH.sub.2Cl.sub.2 to 1% MeOH in
CH.sub.2Cl.sub.2 to 2% MeOH in CH.sub.2Cl.sub.2 to 5% MeOH in
CH.sub.2Cl.sub.2). .sup.1H NMR (CDCl.sub.3) .delta. 7.94 (d, 2H),
7.35-7.23 (m, 5H), 7.11 (m, 1H), 7.00 (d, 2H), 6.93-6.85 (m, 3H),
3.88 (s, 3H), 3.77 (m, 1H), 3.70-3.53 (m, 2H), 2.97 (m, 1H), 2.77
(m, 1H), 2.62 (m, 3H), 2.46-2.26 (m, 2H), 2.06 (m, 1H), 1.96-1.28
(m, 7H).
Step F:
4-(3-{2-[3-Hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-benzoic acid. Analogous to the procedure described for
Compound 1A, Step F,
4-(3-{2-[3-hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-benzoic acid methyl ester was hydrolyzed with 6N NaOH at room
temperature over 24 h to generate
4-(3-{2-[3-hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-benzoic acid. .sup.1H NMR (CDCl.sub.3) .delta. 7.99 (d, 2H),
7.37-7.26 (m, 5H), 7.12 (m, 1H), 7.03-6.88 (m, 5H), 3.82 (m, 1H),
3.66 (m, 2H), 3.00 (m, 1H), 2.85-2.60 (m, 4H), 2.41 (m, 2H), 2.09
(m, 1H), 2.03-1.28 (m, 8H).
Compound 1F
4-{3-[2-(4-Biphenyl-3-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-b-
enzoic acid
Step A: 5-(3-Bromo-3-oxo-butyl)-pyrrolidin-2-one. Analogous to the
procedure described for Compound 1A, Step A,
tetrahydro-pyrrolizine-3,5-dione (5 g, 36 mmol) was reacted with
3-bromobenzylmagnesium bromide (0.25M in Et.sub.2O, 155 mL, 38.8
mmol) over 2 h. Purification by medium pressure chromatography
using a solvent gradient (1:1 hexanes:EtOAc to EtOAc to 5% MeOH in
CH.sub.2Cl.sub.2) provided 5-(3-bromo-3-oxo-butyl)-pyrrolidin-2-one
(7.84 g). .sup.1H NMR (CDCl.sub.3) .delta. 7.41-7.11 (m, 4H), 6.24
(bs, 1H), 3.67 (s, 2H), 3.60 (m, 1H), 2.52 (t, 2H), 2.32 (m, 2H),
2.20 (m, 1H), 1.88-1.60 (m, 3H).
Step B: 5-(3-Bromo-3-hydroxy-butyl)-pyrrolidin-2-one. Analogous to
the procedure described for Compound 1A, Step B,
5-(3-bromo-3-oxo-butyl)-pyrrolidin-2-one (7.84 g, 25.3 mmol) was
reduced with NaBH.sub.4 (480 mg, 12.6 mmol). Purification by medium
pressure chromatography using a solvent gradient (1:1 hexanes:EtOAc
to EtOAc to 1% MeOH in CH.sub.2Cl.sub.2 to 3% MeOH in
CH.sub.2Cl.sub.2 to 5% MeOH in CH.sub.2Cl.sub.2 to 8% MeOH in
CH.sub.2Cl.sub.2) provided
5-(3-bromo-3-hydroxy-butyl)-pyrrolidin-2-one (6.76 g). .sup.1H NMR
(CDCl.sub.3) .delta. 7.36-7.09 (m, 4H), 6.27 (m, 1H), 3.78 (m, 1H),
3.63 (m, 1H), 2.75 (m, 1H), 2.62 (m, 1H), 2.32-2.18 (m, 3H), 1.88
(m, 1H), 1.73-1.42 (m, 5H); MS 312.2, 314.1 (M+).
Step C:
5-[3-Bromo-3-(tert-butyl-dimethyl-silanyloxy)-butyl]-pyrrolidin-2-
-one. Analogous to the procedure described for Compound 1A, Step C,
5-(3-bromo-3-hydroxy-butyl)-pyrrolidin-2-one (6.76 g, 21.6 mmol)
was reacted with tert-butyldimethylsilyl chloride (3.59 g, 23.8
mmol). Purification by medium pressure chromatography using a
solvent gradient (CH.sub.2Cl.sub.2 to 1% MeOH in CH.sub.2Cl.sub.2
to 3% MeOH in CH.sub.2Cl.sub.2 to 5% MeOH in CH.sub.2Cl.sub.2 to 8%
MeOH in CH.sub.2Cl.sub.2) provided
5-[3-bromo-3-(tert-butyl-dimethyl-silanyloxy)-butyl]-pyrrolidin-2-one
(7.45 g). .sup.1H NMR (CDCl.sub.3) .delta. 7.30 (m, 2H), 7.12 (m,
1H), 7.04 (m, 1H), 5.71 (m, 1H), 3.81 (m, 1H), 3.56 (m, 1H), 2.66
(m, 2H), 2.32-2.17 (m, 3H), 1.70-1.35 (m, 5H), 0.82 (s, 9H), -0.06
(d, 3H), -0.24 (d, 3H); MS 426.2, 428.2 (M+).
Step D:
5-[4-Biphenyl-3-yl-3-(tert-butyl-dimethyl-silanyloxy)-butyl]-pyrr-
olidin-2-one. To a solution of
5-[3-bromo-3-(tert-butyl-dimethyl-silanyloxy)-butyl]-pyrrolidin-2-one
(750 mg, 1.76 mmol) in DME (15 mL) was added phenylboronic acid
(236 mg, 1.93 mmol). Palladium acetate (26.8 mg, 0.120 mmol) and
tri-o-tolylphosphine (39.5 mg, 0.130 mmol) were added, followed by
a solution of Na.sub.2CO.sub.3 (373 mg, 3.52 mmol) in water (1.8
mL). The reaction mixture was heated under reflux for 24 h. The
reaction mixture was cooled and the volatiles were removed in
vacuo. The residue was diluted with brine and EtOAc. The aqueous
solution was washed with EtOAc (3.times.) and the combined organic
extracts were dried (MgSO.sub.4), filtered and concentrated.
Purification by medium pressure chromatography eluting with a
solvent gradient (1:1 hexanes:EtOAc to EtOAc to 1% MeOH in
CH.sub.2Cl.sub.2 to 3% MeOH in CH.sub.2Cl.sub.2 to 5% MeOH in
CH.sub.2Cl.sub.2) provided
5-[4-biphenyl-3-yl-3-(tert-butyl-dimethyl-silanyloxy)-butyl]-pyrrolidin-2-
-one (717.3 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.57 (m, 2H),
7.43 (m, 2H), 7.33 (m, 3H), 7.11 (m, 2H), 5.78 (m, 1H), 3.91 (m,
1H), 3.59 (m, 1H), 2.76 (m, 2H), 2.27 (m, 3H), 1.73-1.38 (m, 5H),
0.83 (s, 9H), -0.03 (d, 3H), -0.16 (d, 3H); MS 424.3 (M+1).
Step E:
4-(3-{2-[4-Biphenyl-3-yl-3-(tert-butyl-dimethyl-silanyloxy)-butyl-
]-5-oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester.
Analogous to the procedure described for Compound 1A, Step D,
5-[4-biphenyl-3-yl-3-(tert-butyl-dimethyl-silanyloxy)-butyl]-pyrrolidin-2-
-one (5.116 g, 12.08 mmol) was alkylated with
4-(3-bromo-propyl)-benzoic acid methyl ester (3.41 g, 13.3 mmol)
over 20 h. Purification by medium pressure chromatography using a
solvent gradient (5:1 hexanes:EtOAc to 1:1 hexanes:EtOAc to EtOAc
to 1% MeOH in CH.sub.2Cl.sub.2 to 5% MeOH in CH.sub.2Cl.sub.2)
provided
4-(3-{2-[4-biphenyl-3-yl-3-(tert-butyl-dimethyl-silanyloxy)-butyl]-5-oxo--
pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester (5.38 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.93 (d, 2H), 7.56 (d, 2H), 7.43
(m, 3H), 7.34 (m, 3H), 7.23 (m, 2H), 7.12 (m, 1H), 3.89 (m, 1H),
3.87 (s, 3H), 3.64 (m, 1H), 3.49 (m, 1H), 2.95-2.61 (m, 5H), 2.30
(m, 2H), 2.01 (m, 1H), 1.89-1.70 (m, 3H), 1.59-1.24 (m, 4H), 0.84
(s, 9H), -0.04 (d, 3H), -0.16 (d, 3H).
Step F:
4-{3-[2-(4-Biphenyl-3-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]--
propyl}-benzoic acid methyl ester. Analogous to the procedure
described for Compound 1A, Step E,
4-(3-{2-[4-biphenyl-3-yl-3-(tert-butyl-dimethyl-silanyloxy)-butyl]-5-oxo--
pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester (5.38 g, 8.97
mmol) was deprotected. Purification by medium pressure
chromatography using a solvent gradient (hexanes to 2:1
hexanes:EtOAc to 1:1 hexanes:EtOAc to 0.5% MeOH in CH.sub.2Cl.sub.2
to 1% MeOH in CH.sub.2Cl.sub.2) provided
4-{3-[2-(4-biphenyl-3-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}--
benzoic acid methyl ester (3.70 g). .sup.1H NMR (CDCl.sub.3)
.delta. 7.93 (d, 2H), 7.57 (d, 2H), 7.40 (m, 6H), 7.24 (m, 2H),
7.17 (m, 1H), 3.86 (s, 3H), 3.80 (m, 1H), 3.66 (m, 1H), 3.56 (m,
1H), 2.97 (m, 1H), 2.90-2.60 (m, 4H), 2.33 (m, 2H), 2.07 (m, 1H),
1.98-1.34 (m, 8H).
Step G:
4-{3-[2-(4-Biphenyl-3-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]--
propyl}-benzoic acid. Analogous to the procedure described for
Compound 1A, Step F,
4-{3-[2-(4-biphenyl-3-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}--
benzoic acid methyl ester (3.14 g, 6.47 mmol) was hydrolyzed with
6N NaOH (40 mL) in MeOH (160 mL) at room temperature over 24 h to
generate
4-{3-[2-(4-biphenyl-3-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}--
benzoic acid (2.73 g). .sup.1H NMR (CDCl.sub.3) .delta. 7.98 (d,
2H), 7.57 (d, 2H), 7.40 (m, 6H), 7.26 (m, 2H), 7.18 (m, 1H), 3.85
(m, 1H), 3.68 (m, 1H), 3.59 (m, 1H), 2.98 (m, 1H), 2.88 (m, 1H),
2.70 (m, 3H), 2.36 (m, 2H), 2.08 (m, 1H), 1.85 (m, 3H), 1.69-1.35
(m, 4H); MS 470.1 (M-1), 472.2 (M+1).
Compound 1G
4-(3-{2-[4-(4-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-propy-
l)-benzoic acid
Step A: 5-[4-(4-Fluoro-phenyl)-3-oxo-butyl]-pyrrolidin-2-one.
Analogous to the procedure described for Compound 1A, Step A,
tetrahydro-pyrrolizine-3,5-dione (1.41 g, 10.1 mmol) was reacted
with 4-fluorobenzylmagnesium chloride (0.25M in Et.sub.2O, 50 mL,
12.5 mmol) over 5 h. Purification by medium pressure chromatography
(2% MeOH in CH.sub.2Cl.sub.2) provided
5-[4-(4-fluoro-phenyl)-3-oxo-butyl]-pyrrolidin-2-one (2.64 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.18 (m, 2H), 7.03 (m, 2H), 6.34
(m, 1H), 3.70 (s, 2H), 3.62 (m, 1H), 2.54 (t, 2H), 2.34-2.15 (m,
3H), 1.82-1.61 (m, 3H).
Step B: 5-[4-(4-Fluoro-phenyl)-3-hydroxy-butyl]-pyrrolidin-2-one.
Analogous to the procedure described for Compound 1A, Step B,
5-[4-(4-fluoro-phenyl)-3-oxo-butyl]-pyrrolidin-2-one (2.64 g, 10.6
mmol) was reduced with NaBH.sub.4 (400 mg, 10.5 mmol) at room
temperature for 1 h. Additional NaBH.sub.4 (150 mg, 3.95 mmol) was
added and the reaction mixture was stirred for 20 h. Purification
by medium pressure chromatography using a solvent gradient
(CH.sub.2Cl.sub.2 to 2% MeOH in CH.sub.2Cl.sub.2 to 4% MeOH in
CH.sub.2Cl.sub.2) provided
5-[4-(4-fluoro-phenyl)-3-hydroxy-butyl]-pyrrolidin-2-one (2.01 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.14 (m, 2H), 6.98 (m, 2H), 6.78
(m, 1H), 3.76 (m, 1H), 3.65 (m, 1H), 2.76 (m, 1H), 2.64 (m, 1H),
2.32-2.18 (m, 4H), 1.72-1.47 (m, 5H).
Step C:
5-[3-(tert-Butyl-dimethyl-silanyloxy)-4-(4-fluoro-phenyl)-butyl]--
pyrrolidin-2-one. Analogous to the procedure described for Compound
1A, Step C,
5-[4-(4-fluoro-phenyl)-3-hydroxy-butyl]-pyrrolidin-2-one (1.95 g,
7.79 mmol) was reacted with tert-butyldimethylsilyl chloride (1.47
g, 9.76 mmol). Purification by medium pressure chromatography (1%
MeOH in CH.sub.2Cl.sub.2) provided
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-(4-fluoro-phenyl)-butyl]-pyrrolid-
in-2-one. .sup.1H NMR (CDCl.sub.3) .delta. 7.12 (m, 2H), 6.97 (m,
2H), 5.75 (m, 1H), 3.83 (m, 1H), 3.60 (m, 1H), 2.71 (m, 2H),
2.36-2.24 (m, 3H), 1.70-1.38 (m, 5H), 0.84 (s, 9H), -0.05 (d, 3H),
-0.2 (d, 3H).
Step D:
4-(3-{2-[3-(tert-Butyl-dimethyl-silanyloxy)-4-(4-fluoro-phenyl)-b-
utyl]-5-oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester.
Analogous to the procedure described for Compound 1A, Step D,
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-(4-fluoro-phenyl)-butyl]-pyrrolid-
in-2-one (296 mg, 0.809 mmol) was alkylated with
4-(3-bromo-propyl)-benzoic acid methyl ester (276 mg, 1.07 mmol)
over 72 h. Purification by medium pressure chromatography (1:1
hexanes:EtOAc) provided
4-(3-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-(4-fluoro-phenyl)--
butyl]-5-oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester
(250 mg). .sup.1H NMR (CDCl.sub.3) (selected peaks) .delta. 7.92
(d, 2H), 7.21 (d, 2H), 7.05 (m, 2H), 6.92 (m, 2H), 3.86 (s, 3H),
3.76 (m, 1H), 3.62 (m, 1H), 3.45 (m, 1H), 0.81 (s, 9H).
Step E:
4-(3-{2-[4-(4-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1--
yl}-propyl)-benzoic acid methyl ester. Analogous to the procedure
described for Compound 1A, Step E,
4-(3-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-(4-fluoro-phenyl)-butyl]-5--
oxo-pyrrolidin-1-yl}-propyl)-benzoic acid methyl ester (241.2 mg,
0.445 mmol) was deprotected to yield, after medium pressure
chromatography (1:1 hexanes:EtOAc to EtOAc to 1% MeOH in
CH.sub.2Cl.sub.2 to 3% MeOH in CH.sub.2Cl.sub.2 to 5% MeOH in
CH.sub.2Cl.sub.2),
4-(3-{2-[4-(4-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-benzoic acid methyl ester (61.1 mg). .sup.1H NMR (CDCl.sub.3)
(selected peaks) .delta. 7.93 (d, 2H), 7.24 (d, 2H), 7.14 (m, 2H),
7.00 (m, 2H), 3.88 (s, 3H), 3.80-3.51 (m, 3H), 2.98 (m, 1H), 2.32
(m, 2H).
Step F:
4-(3-{2-[4-(4-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1--
yl}-propyl)-benzoic acid. Analogous to the procedure described for
Compound 1A, Step F,
4-(3-{2-[4-(4-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-benzoic acid methyl ester (61.1 mg, 0.143 mmol) was hydrolyzed
with 6N NaOH (1 mL) in MeOH (5 mL) at room temperature over 24 h.
Purification by medium pressure chromatography eluting with a
solvent gradient (CH.sub.2Cl.sub.2 to 2% MeOH in CH.sub.2Cl.sub.2
to 4% MeOH in CH.sub.2Cl.sub.2 to 6% MeOH in CH.sub.2Cl.sub.2 to
10% MeOH in CH.sub.2Cl.sub.2) provided the title compound (45 mg).
.sup.1H NMR (CDCl.sub.3) .delta. 7.97 (d, 2H), 7.25 (m, 2H), 7.14
(m, 2H), 6.99 (m, 2H), 3.75-3.58 (m, 3H), 2.97 (m, 1H), 2.69 (m,
4H), 2.40 (m, 2H), 2.15-1.35 (m, 9H); MS 413.8 (M+).
Compound 1H
4-{2-[2-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-ethoxy}-benzoic
acid
Step A:
4-(2-{2-[3-(tert-Butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-5-oxo-
-pyrrolidin-1-yl}-ethoxy)-benzoic acid ethyl ester. Analogous to
the procedure described for Compound 1A, Step D,
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-pyrrolidin-2-one
(prepared in Compound 1A, Step C) (250 mg, 0.719 mmol) was
alkylated with NaHMDS (1M in THF, 0.86 mL, 0.86 mmol) and
4-(2-bromo-ethoxy)-benzoic acid ethyl ester (216 mg, 0.791 mmol).
The reaction temperature was maintained at 50.degree. C. over 24 h.
Purification by radial chromatography (hexanes to 4:1
hexanes:EtOAc) provided
4-(2-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-5-oxo-pyrroli-
din-1-yl}-ethoxy)-benzoic acid ethyl ester (66.4 mg). .sup.1H NMR
(CDCl.sub.3) (selected peaks) .delta. 7.96 (m, 2H), 7.29-7.13 (m,
5H), 6.84 (m, 2H), 4.33 (q, 2H), 4.12 (m, 2H), 3.90 (m, 2H), 3.68
(m, 1H), 3.34 (m, 1H), 2.73 (m, 2H), 2.32 (m, 2H), 1.36 (t, 3H),
0.85 (s, 9H), -0.03 (s, 3H), -0.15 (d, 3H).
Step B:
4-{2-[2-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-ethoxyl-
}-benzoic acid ethyl ester. Analogous to the procedure described
for Compound 1A, Step E,
4-(2-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-5-oxo-pyrroli-
din-1-yl}-ethoxy)-benzoic acid ethyl ester (66.4 mg, 0.122 mmol)
was deprotected to provide
4-{2-[2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-ethoxy}-benzoic
acid ethyl ester (52 mg) after purification by radial
chromatography (CH.sub.2Cl.sub.2 to 2% MeOH in CH.sub.2Cl.sub.2).
.sup.1H NMR (CDCl.sub.3) .delta. 7.94 (m, 2H), 7.31-7.16 (m, 5H),
6.83 (m, 2H), 4.30 (q, 2H), 4.12 (m, 2H), 3.90 (m, 1H), 3.76 (m,
2H), 3.38 (m, 1H), 2.80 (m, 1H), 2.64 (m, 1H), 2.33 (m, 2H), 2.10
(m, 1H), 1.69-1.37 (m, 6H), 1.34 (t, 3H).
Step C:
4-{2-[2-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-ethoxy}-
-benzoic acid. Analogous to the procedure described for Compound
1A, Step F,
4-{2-[2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-ethoxy}-benz-
oic acid ethyl ester (52 mg, 0.122 mmol) was hydrolyzed with 6N
NaOH (1 mL) to yield the title compound (41.5 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.98 (d, 2H), 7.32-7.16 (m, 5H), 6.85 (m, 2H),
4.13 (m, 2H), 3.92 (m, 1H), 3.81 (m, 1H), 3.75 (m, 1H), 3.40 (m,
1H), 2.82 (m, 1H), 2.66 (m, 1H), 2.36 (m, 2H), 2.10 (m, 2H),
1.70-1.34 (m, 5H); MS 398.4 (M+1), 396.3 (M-1).
Compound 2A
7-{2S-[3R-Hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl}-
-heptanoic acid
Step A: 7-(2R-Formyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid ethyl
ester. To a solution of
7-(2R-hydroxymethyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid ethyl
ester (1.63 g, 6.01 mmol) in anhydrous benzene (50 mL) was added
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.46
g, 18.03 mmol) and DMSO (1.5 mL, 24.04 mmol). The solution was
cooled to 0.degree. C. and pyridinium trifluoroacetate (1.28 g,
6.61 mmol) was added. The reaction mixture was stirred at 0.degree.
C. for 15 minutes and at room temperature for 2 h. The solution was
decanted from the oily residue. The residue was washed with benzene
(3.times.) and the combined benzene washes were concentrated in
vacuo to provide 7-(2R-formyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid
ethyl ester, which was used in Step B without further
purification.
Step B:
7-{2R-[4-(3-Methoxymethyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrroli-
din-1-yl}-heptanoic acid ethyl ester. To a solution of
[3-(3-methoxymethyl-phenyl)-2-oxo-propyl]-phosphonic acid diethyl
ester (1.715 g, 5.46 mmol) in THF (43 mL) at 0.degree. C. was added
NaH (60% by weight in oil, 240 mg, 6.00 mmol) portionwise. The
reaction mixture was stirred at room temperature for 45 minutes.
The reaction mixture was cooled to 0.degree. C. and a solution of
7-(2R-formyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid ethyl ester
(prepared in Step A, assumed 6.01 mmol) in THF (32 mL) was added
dropwise. The reaction mixture was stirred at 0.degree. C. for 15
minutes and at room temperature for 24 h. The reaction mixture was
cooled to 0.degree. C. and acetic acid was added until a pH of 5
was achieved. EtOAc and water were added and the aqueous solution
was washed with EtOAc (3.times.). The organic solutions were
combined, washed with water, dried (MgSO.sub.4), filtered and
concentrated. The residue was purified by medium pressure
chromatography eluting with a solvent gradient (2:1 hexanes:EtOAc
to 1:1 hexanes:EtOAc to 1% MeOH in CH.sub.2Cl.sub.2 to 3% MeOH in
CH.sub.2Cl.sub.2) to provide
7-{2R-[4-(3-methoxymethyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl-
}-heptanoic acid ethyl ester (1.4 g). .sup.1H NMR (CDCl.sub.3)
.delta. 7.29 (m, 1H), 7.22 (m, 1H), 7.16 (s, 1H), 7.09 (d, 1H),
6.62 (dd, 1H), 6.19 (d, 1H), 4.41 (s, 2H), 4.10 (m, 3H), 3.82 (s,
2H), 3.51 (m, 1H), 3.36 (s, 3H), 2.67 (m, 1H), 2.43-2.18 (m, 5H),
1.75 (m, 1H), 1.56 (m, 2H), 1.42-1.17 (m, 9H).
Step C:
7-{2R-[3S-Hydroxy-4-(3-methoxymethyl-phenyl)-but-1-enyl]-5-oxo-py-
rrolidin-1-yl}-heptanoic acid ethyl ester. To a solution of
7-{2R-[4-(3-methoxymethyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl-
}-heptanoic acid ethyl ester (1.40 g, 3.26 mmol) in anhydrous
CH.sub.2Cl.sub.2 (200 mL) was added
(R)-2-methyl-CBS-oxazaborolidine (1M in toluene, 0.49 mL, 0.49
mmol) and the solution was cooled to -45.degree. C. The reaction
mixture was stirred for 20 minutes and catecholborane (1M in THF,
9.8 mL, 9.8 mmol) was added. The reaction mixture was stirred for
24 h at -45.degree. C. and THF (100 mL) and HCl (1N, 100 mL) were
added. The reaction mixture was stirred at room temperature for 24
h and at 40-45.degree. C. for 1.5 h. The solution was diluted with
CH.sub.2Cl.sub.2 and water and the layers were separated. The
organic solution was cooled to 0.degree. C. and was washed with
ice-cold NaOH (0.5N) followed by brine. The organic solution was
again washed with ice-cold NaOH (0.5 N) followed by brine and was
dried (MgSO.sub.4), filtered and concentrated. Purification by
medium pressure chromatography eluting with a solvent gradient (5:1
hexanes:EtOAc to 2:1 hexanes:EtOAc to 1:1 hexanes:EtOAc to EtOAc to
2% MeOH in CH.sub.2Cl.sub.2) provided
7-{2R-[3S-hydroxy-4-(3-methoxymethyl-phenyl)-but-1-enyl]-5-oxo-pyrrolidin-
-1-yl}-heptanoic acid ethyl ester (1.2 g) as an approximate 12:1
mixture of 3S:3R alcohol diasteromers by HPLC analysis. .sup.1H NMR
(CDCl.sub.3) (selected peaks) .delta. 7.26-7.07 (m, 4H), 5.67 (m,
1H), 5.43 (m, 1H), 4.39 (s, 2H), 4.36 (m, 1H), 4.06 (q, 2H), 3.98
(m, 1H), 3.41 (m, 1H), 3.35 (s, 3H); MS 432.3 (M+1), 430.3
(M-1).
Step D:
7-{2S-[3R-Hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-5-oxo-pyrroli-
din-1-yl}-heptanoic acid ethyl ester. To a solution of
7-{2R-[3S-hydroxy-4-(3-methoxymethyl-phenyl)-but-1-enyl]-5-oxo-pyrrolidin-
-1-yl}-heptanoic acid ethyl ester (1.2 g, 2.78 mmol) in EtOH (100
mL) was added 10% palladium on carbon (120 mg). The reaction
mixture was hydrogenated on a Parr shaker at 45 psi for 24 h. The
catalyst was removed via filtration through Celite.RTM. with the
aid of EtOH. Purification by medium pressure chromatography eluting
with a solvent gradient (CH.sub.2Cl.sub.2 to 2% MeOH in
CH.sub.2Cl.sub.2 to 5% MeOH in CH.sub.2Cl.sub.2) (2.times.)
provided
7-{2S-[3R-hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl-
}-heptanoic acid ethyl ester (1.1 g). .sup.1H NMR (CDCl.sub.3)
.delta. 7.28 (m, 1H), 7.18 (m, 2H), 7.11 (m, 1H), 4.42 (s, 2H),
4.08 (q, 2H), 3.82 (m, 1H), 3.58 (m, 2H), 3.38 (s, 3H), 2.84 (m,
2H), 2.66 (m, 1H), 2.41-2.23 (m, 4H), 2.08 (m, 1H), 1.78 (m, 1H),
1.64-1.37 (m, 9H), 1.28 (m, 4H), 1.22 (t, 3H).
Step E:
7-{2S-[3R-Hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-5-oxo-pyrroli-
din-1-yl}-heptanoic acid. To a solution of
7-{2S-[3R-hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl-
}-heptanoic acid ethyl ester (1.1 g, 2.53 mmol) in EtOH (32 mL) was
added NaOH (6N, 16 mL). The reaction mixture was stirred for 24 h
and 1N HCl was added to obtain a pH of about 2. Brine and
CH.sub.2Cl.sub.2 were added and the layers were separated. The
aqueous solution was washed with 5% MeOH in CH.sub.2Cl.sub.2 (2
times). The combined organic layers were dried (MgSO.sub.4),
filtered and concentrated to provide the title compound of Example
2A (990 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.28 (m, 1H), 7.18
(m, 2H), 7.11 (m, 1H), 4.43 (s, 2H), 3.83 (m, 1H), 3.57 (m, 2H),
3.40 (s, 3H), 2.91 (m, 1H), 2.79 (m, 1H), 2.66 (m, 1H), 2.43-2.25
(m, 4H), 2.10 (m, 1H), 1.83 (m, 1H), 1.66-1.22 (m, 13H); MS 406.3
(M+1), 404.3 (M-1).
Compound 2B
7-[2R-(3-Hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-heptanoic
acid
Step A:
7-[2R-(4-Naphthalen-2-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-
-heptanoic acid ethyl ester. Analogous to the procedure described
for Compound 2A, Step B, the anion derived from
(3-naphthalen-2-yl-2-oxo-propyl)-phosphonic acid dimethyl ester
(646 mg, 2.21 mmol) and NaH (60% by weight in oil, 81 mg, 2.02
mmol) was reacted with
7-(2R-formyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid ethyl ester
(assumed 1.84 mmol) over 163 h. Purification by medium pressure
chromatography (1:1 hexanes:EtOAc to EtOAc) provided
7-[2R-(4-naphthalen-2-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-heptano-
ic acid ethyl ester (340 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.78
(m, 3H), 7.65 (s, 1H), 7.46 (m, 2H), 7.30 (d, 1H), 6.66 (dd, 1H),
6.24 (d, 1H), 4.10 (m, 3H), 3.99 (s, 2H), 3.45 (m, 1H), 2.63 (m,
1H), 2.44-2.18 (m, 5H), 1.75 (m, 1H), 1.52 (m, 2H), 1.37-1.06 (m,
9H); MS 436.1 (M+1), 434.1 (M-1).
Step B:
7-[2S-(4-Naphthalen-2-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-hept-
anoic acid ethyl ester. Analogous to the procedure described for
Compound 2A, Step D, a mixture of
7-[2R-(4-naphthalen-2-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-heptano-
ic acid ethyl ester (337 mg, 0.774 mmol) and 10% palladium on
carbon (50 mg) in EtOH (50 mL) was hydrogenated at 50 psi for 3 h.
Medium pressure chromatography (1:1 hexanes:EtOAc to EtOAc)
provided
7-[2S-(4-naphthalen-2-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-heptanoic
acid ethyl ester (290 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.80
(m, 3H), 7.66 (s, 1H), 7.47 (m, 2H), 7.30 (m, 1H), 4.10 (q, 2H),
3.85 (s, 2H), 3.52 (m, 2H), 2.77 (m, 1H), 2.47 (m, 2H), 2.26 (m,
4H), 1.98 (m, 2H), 1.61-1.16 (m, 13H); MS 438.1 (M+1), 436.1
(M-1).
Step C:
7-[2S-(3-Hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]--
heptanoic acid ethyl ester. To a solution of
7-[2S-(4-naphthalen-2-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-heptanoic
acid ethyl ester (367 mg, 0.839 mmol) in EtOH (20 mL) was added
NaBH.sub.4 (32 mg, 0.839 mmol). The reaction mixture was stirred
for 2 h and water (5 mL) was added. The volatiles were removed in
vacuo and the remaining aqueous solution was washed with CHCl.sub.3
(4.times.10 mL). The organic solutions were combined, dried
(MgSO.sub.4), filtered and concentrated. Purification by medium
pressure chromatography (1:1 hexanes:EtOAc to EtOAc) provided
7-[2S-(3-hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-heptanoi-
c acid ethyl ester (332 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.80
(m, 3H), 7.65 (s, 1H), 7.46 (m, 2H), 7.33 (m, 1H), 4.07 (m, 2H),
3.91 (m, 1H), 3.60 (m, 2H), 2.98 (m, 1H), 2.84 (m, 2H), 2.35 (m,
2H), 2.25 (t, 2H), 2.10 (m, 1H), 2.01 (m, 1H), 1.81 (m, 1H), 1.70
(d, 1H), 1.68-1.37 (m, 7H), 1.36-1.20 (m, 7H); MS 440.1 (M+1).
Step D:
7-[2S-(3-Hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]--
heptanoic acid. A solution of
7-[2S-(3-hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-heptanoi-
c acid ethyl ester (327 mg, 0.744 mmol), NaOH (1M, 0.8 mL), and
MeOH (15 mL) was heated under reflux for 4 h. The volatiles were
removed in vacuo and water (15 mL) was added. The aqueous solution
was acidified to a pH of 5 with 1N HCl and the acidic solution was
washed with CHCl.sub.3 (4.times.10 mL). The organic solutions were
combined, dried (MgSO.sub.4), filtered and concentrated to provide
7-[2S-(3-hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-heptanoi-
c acid (180 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.80 (m, 3H),
7.65 (s, 1H), 7.46 (m, 2H), 7.33 (m, 1H), 3.94 (m, 1H), 3.58 (m,
2H), 3.02-2.80 (m, 3H), 2.34 (m, 4H), 2.08 (m, 2H), 1.67-1.23 (m,
13H); MS 412.1 (M+1), 410.2 (M-1).
Step E: Sodium salt of
7-[2S-(3-hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-heptanoi-
c acid. To a solution of
7-[2S-(3-hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-heptanoi-
c acid (35 mg, 0.0851 mmol) in MeOH (5 mL) at 0.degree. C. was
added NaOH (1M, 0.085 mL). The reaction mixture was stirred for 1.5
h at 0.degree. C. and was concentrated in vacuo, azeotroping with
CHCl.sub.3 (3.times.5 mL) to yield the sodium salt of the title
compound of Example 2B (37 mg). .sup.1H NMR (CDCl.sub.3) .delta.
7.69-7.24 (m, 7H), 3.78 (m, 1H), 3.40 (m, 2H), 2.80 (m, 6H),
2.16-1.70 (m, 4H), 1.43-1.18 (m, 12H).
Compound 2C
7-[2R-(4-Benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrolidin-1-yl-
]-heptanoic acid
Step A:
7-[2R-(4-Benzo[1,3]dioxol-5-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-
-1-yl]-heptanoic acid ethyl ester. Analogous to the procedure
described for Compound 2A, Step B, the anion generated from
(3-benzo[1,3]dioxol-5-yl-2-oxo-propyl)-phosphonic acid dimethyl
ester (12.65 g, 44.2 mmol) and NaH (60% by weight in oil, 1.62 g,
40.5 mmol) was reacted with
7-(2R-formyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid ethyl ester
(assumed 36.8 mmol) over 24 h. Purification by medium pressure
chromatography (10% EtOAc in hexanes to 40% EtOAc in hexanes)
provided
7-[2R-(4-benzo[1,3]dioxol-5-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-h-
eptanoic acid ethyl ester (4.18 g). .sup.1H NMR (CDCl.sub.3)
.delta. 6.76 (d, 1H), 6.63 (m, 3H), 6.20 (d, 1H), 5.94 (s, 2H),
4.13 (m, 3H), 3.74 (s, 2H), 3.52 (m, 1H), 2.71 (m, 1H), 2.38 (m,
2H), 2.26 (m, 3H), 1.78 (m, 1H), 1.58 (m, 5H), 1.46-1.19 (m,
6H).
Step B:
7-[2R-(4-Benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrol-
idin-1-yl]-heptanoic acid ethyl ester. Analogous to the procedure
described for Compound 2B, Step C,
7-[2R-(4-benzo[1,3]dioxol-5-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-h-
eptanoic acid ethyl ester (4.18 g, 9.74 mmol) was reacted with
NaBH.sub.4 (369 mg, 9.74 mmol) in EtOH (32 mL). The NaBH.sub.4
addition was performed at 0.degree. C. and the reaction mixture was
stirred at room temperature for 3 h. Purification by medium
pressure chromatography (EtOAc) provided
7-[2R-(4-benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrolidin-1-y-
l]-heptanoic acid ethyl ester (3.36 g).
Step C:
7-[2R-(4-Benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrol-
idin-1-yl]-heptanoic acid. Analogous to the procedure described for
Compound 2A, Step E,
7-[2R-(4-benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrolidin-1-y-
l]-heptanoic acid ethyl ester (3.36 g, 7.79 mmol) was hydrolyzed
with 2N NaOH (11 mL) in MeOH. Purification by medium pressure
chromatography (50% EtOAc in hexanes to EtOAc to 5% MeOH in
CH.sub.2Cl.sub.2) followed by a second column eluting with a
solvent gradient (1% MeOH to CH.sub.2Cl.sub.2 to 5% MeOH in
CH.sub.2Cl.sub.2) provided
7-[2R-(4-benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrolidin-1-y-
l]-heptanoic acid (2.26 g). .sup.1H NMR (CDCl.sub.3) .delta. 6.66
(m, 3H), 5.91 (s, 2H), 5.69 (m, 1H), 5.44 (m, 1H), 4.31 (m, 1H),
4.01 (m, 1H), 3.45 (m, 1H), 2.76 (m, 3H), 2.34 (m, 4H), 2.15 (m,
1H). 1.70-1.20 (m, 10H); MS 404.3 (M+1), 402.1 (M-1).
Step D: Sodium salt of
7-[2R-(4-Benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrolidin-1-y-
l]-heptanoic acid. The sodium salt was prepared by addition of
NaHCO.sub.3 (470 mg, 5.60 mmol) in water to a solution of
7-[2R-(4-benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrolidin-1-y-
l]-heptanoic acid (2.26 g, 5.60 mmol) in EtOH. The reaction mixture
was stirred for 3 h and was concentrated in vacuo to provide the
sodium salt of the title compound, Compound 2C. .sup.1H NMR
(CD.sub.3OD) .delta. 6.65 (m, 3H), 5.85 (s, 2H), 5.67 (m, 1H), 5.34
(m, 1H), 4.24 (m, 1H), 4.09 (m, 1H), 3.45 (m, 1H), 2.79 (m, 2H),
2.61 (m, 2H), 2.29 (m, 2H), 2.16 (m, 3H), 1.68-1.17 (m, 9H).
Compound 2D
7-[2S-(4-Benzo[1,3]dioxol-5-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]-hep-
tanoic acid
Step A:
7-[2S-(4-Benzo[1,3]dioxol-5-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin--
1-yl]-heptanoic acid. Analogous to the procedure described for
Compound 2A, Step D, a mixture of
7-[2R-(4-benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrolidin-1-y-
l]-heptanoic acid (120 mg, 2.96 mmol), MeOH (30 mL), and 10%
palladium on carbon (14 mg) was hydrogenated at 50 psi for 18 h to
provide
7-[2S-(4-benzo[1,3]dioxol-5-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]-he-
ptanoic acid (71.3 mg). .sup.1H NMR (CDCl.sub.3) .delta. 6.68 (m,
3H), 5.92 (s, 2H), 3.74 (m, 1H), 3.57 (m, 2H), 2.87 (m, 1H), 2.72
(m, 1H), 2.54 (m, 1H), 2.31 (m, 4H), 2.10 (m, 1H), 1.99 (m, 1H),
1.66-1.19 (m, 13H); MS 406.3 (M+1), 404.3 (M-1).
Compound 2E
4-{3-[2R-(4-Benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrolidin-1-
-yl]-propyl}-benzoic acid
Step A:
4-{3-[2R-(4-Benzo[1,3]dioxol-5-yl-3-oxo-but-1-enyl)-5-oxo-pyrroli-
din-1-yl]-propyl}-benzoic acid methyl ester. Analogous to the
procedure described for Compound 2A, Step B, the anion derived from
(3-benzo[1,3]dioxol-5-yl-2-oxo-propyl)-phosphonic acid dimethyl
ester (356 mg, 1.28 mmol) and NaH (60% in oil, 46 mg, 1.14 mmol)
was reacted with
4-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-benzoic acid methyl
ester (assumed 1.04 mmol) over 24 h. Purification by medium
pressure chromatography (30% hexane in EtOAc to EtOAc) provided
4-{3-[2R-(4-benzo[1,3]dioxol-5-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl-
]-propyl}-benzoic acid methyl ester (202 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.92 (d, 2H), 7.18 (d, 2H), 6.73 (d, 1H), 6.60
(m, 3H), 6.15 (d, 1H), 5.91 (s, 2H), 4.08 (m, 1H), 3.87 (s, 3H),
3.68 (s, 2H), 3.56 (m, 1H), 2.79 (m, 1H), 2.59 (t, 2H), 2.34 (m,
2H), 2.14 (m, 1H), 1.72 (m, 3H); MS 450.1 (M+1).
Step B:
4-{3-[2R-(4-Benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyr-
rolidin-1-yl]-propyl}-benzoic acid methyl ester. Analogous to the
procedure described for Compound 2B, Step C,
4-{3-[2R-(4-benzo[1,3]dioxol-5-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl-
]-propyl}-benzoic acid methyl ester (202 mg, 0.449 mmol) was
reacted with NaBH.sub.4 (17 mg, 0.45 mmol) in MeOH (8 mL) at
0.degree. C. over 2 h. Purification by medium pressure
chromatography (EtOAc to 2% MeOH in CH.sub.2Cl.sub.2) provided
4-{3-[2R-(4-benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrolidin--
1-yl]-propyl}-benzoic acid methyl ester (156 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.94 (d, 2H), 7.23 (d, 2H), 6.67 (m, 3H), 5.92
(s, 2H), 5.66 (m, 1H), 5.45 (m, 1H), 4.28 (m, 1H), 3.99 (m, 1H),
3.87 (s, 3H), 3.55 (m, 1H), 2.88-2.59 (m, 5H), 2.50-1.61 (m, 7H);
MS 452.1 (M+1).
Step C:
4-{3-[2R-(4-Benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyr-
rolidin-1-yl]-propyl}-benzoic acid. Analogous to the procedure
described for Compound 2A, Step E,
4-{3-[2R-(4-benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrolidin--
1-yl]-propyl}-benzoic acid methyl ester (156 mg, 0.345 mmol) was
hydrolyzed with 2N NaOH in MeOH (5 mL) to provide the title
compound of Example 2E (120 mg). .sup.1H NMR (CDCl.sub.3) .delta.
7.99 (d, 2H), 7.26 (m, 2H), 6.74 (d, 1H), 6.63 (m, 2H), 5.91 (s,
2H), 5.67 (m 1H), 5.46 (m, 1H), 4.29 (m, 1H), 3.99 (m, 1H), 3.57
(m, 1H), 2.94-2.60 (m, 5H), 2.36 (m, 2H), 2.14 (m, 1H), 1.87-1.62
(m, 4H); MS 436.2 (M-1).
Compound 2F
4-{3-[2S-(4-Benzo[1,3]dioxol-5-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]--
propyl}-benzoic acid
Step A:
4-{3-[2S-(4-Benzo[1,3]dioxol-5-yl-3-hydroxy-butyl)-5-oxo-pyrrolid-
in-1-yl]-propyl}-benzoic acid. Analogous to the procedure described
for Compound 2A, Step D,
4-{3-[2R-(4-benzo[1,3]dioxol-5-yl-3-hydroxy-but-1-enyl)-5-oxo-pyrrolidin--
1-yl]-propyl}-benzoic acid (116 mg, 0.265 mmol) was hydrogenated to
provide
4-{3-[2S-(4-benzo[1,3]dioxol-5-yl-3-hydroxy-butyl)-5-oxo-pyrrolid-
in-1-yl]-propyl}-benzoic acid (101 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 7.99 (d, 2H), 7.26 (m, 2H), 6.74 (d, 1H), 6.63 (m, 2H),
5.91 (s, 2H), 5.68 (m, 1H), 5.46 (m, 1H), 4.29 (m, 1H), 3.99 (m,
1H), 3.56 (m, 1H), 2.91 (m, 4H), 2.84-2.60 (m, 4H), 2.36 (m, 2H),
2.14 (m, 1H), 1.87-1.62 (m, 4H); MS 438.2 (M-1).
Compound 2G
7-{2S-[3R-Hydroxy-4-(3-trifluoromethoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1--
yl}-heptanoic acid
Step A:
7-{2-Oxo-5R-[3-oxo-4-(3-trifluoromethoxy-phenyl)-but-1-enyl]-pyrr-
olidin-1-yl}-heptanoic acid ethyl ester. Analogous to the procedure
described for Compound 2A, Step B, the anion derived from
[2-oxo-3-(3-trifluoromethoxy-phenyl)-propyl]-phosphonic acid
dimethyl ester (370 mg, 1.13 mmol) and NaH (60% in oil, 45 mg, 1.13
mmol) was reacted with
7-(2R-formyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid ethyl ester
(assumed 1.13 mmol) over 16 h. Medium pressure chromatography (19:1
hexanes:EtOAc to 6:4 hexanes:EtOAc to 1:1 hexanes:EtOAc to EtOAc)
provided
7-{2-oxo-5R-[3-oxo-4-(3-trifluoromethoxy-phenyl)-but-1-enyl]-pyr-
rolidin-1-yl}-heptanoic acid ethyl ester (132 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.35 (m, 1H), 7.12 (m, 2H), 7.05 (s, 1H), 6.66
(dd, 1H), 6.21 (d, 1H), 4.18 (m, 1H), 4.10 (q, 2H), 3.86 (s, 2H),
3.54 (m, 1H), 2.70 (m, 1H), 2.47-2.22 (m, 5H), 1.78 (m, 1H), 1.57
(m, 2H), 1.61-1.21 (m, 9H); MS 470.2 (M+1), 468.1 (M-1).
Step B:
7-{2R-[3S-Hydroxy-4-(3-trifluoromethoxy-phenyl)-but-1-enyl]-5-oxo-
-pyrrolidin-1-yl}-heptanoic acid ethyl ester. To a solution of
7-{2-oxo-5R-[3-oxo-4-(3-trifluoromethoxy-phenyl)-but-1-enyl]-pyrrolidin-1-
-yl}-heptanoic acid ethyl ester (169 mg, 0.360 mmol) and
(R)-2-methyl-CBS-oxazaborolidine (1M in toluene, 0.054 mL, 0.054
mmol) in CH.sub.2Cl.sub.2 (25.0 mL) at -45.degree. C. was added
catecholborane (1M in THF, 1.08 mL, 1.08 mmol) dropwise. The
reaction mixture was stirred at -45.degree. C. for 19 h. methanol
(5 mL) was added and the reaction mixture was warmed to room
temperature and was concentrated in vacuo. The residue was
dissolved in CHCl.sub.3 and the organic solution was washed with 1M
NaOH (4.times.10 mL), 1M HCl (1.times.10 mL), and water (1.times.10
mL). The organic solution was dried (MgSO.sub.4), filtered and
concentrated. Purification by medium pressure chromatography (9:1
hexanes:EtOAc to 1:1 hexanes:EtOAc to EtOAc) provided
7-{2R-[3S-hydroxy-4-(3-trifluoromethoxy-phenyl)-but-1-enyl]-5-oxo-pyrroli-
din-1-yl}-heptanoic acid ethyl ester (90 mg) as a 9:1 mixture
(3S:3R) of alcohol diastereomers by HPLC analysis. .sup.1H NMR
(CDCl.sub.3) .delta. 7.32 (m, 1H), 7.10 (m, 3H), 5.70 (dd, 1H),
5.50 (dd, 1H), 4.41 (m, 1H), 4.09 (q, 2H), 4.01 (m, 1H), 3.45 (m,
1H), 2.85 (d, 2H), 2.70 (m, 1H), 2.41-2.24 (m, 4H), 2.17 (m, 1H),
1.71-1.54 (m, 5H), 1.47-1.21 (m, 8H); MS 472.3 (M+1), 470.2
(M-1).
Step C:
7-{2S-[3R-Hydroxy-4-(3-trifluoromethoxy-phenyl)-butyl]-5-oxo-pyrr-
olidin-1-yl}-heptanoic acid ethyl ester. Analogous to the procedure
described for Compound 2A, Step D, a solution of
7-{2R-[3S-hydroxy-4-(3-trifluoromethoxy-phenyl)-but-1-enyl]-5-oxo-pyrroli-
din-1-yl}-heptanoic acid ethyl ester (86 mg, 0.182 mmol) in EtOH
(40 mL) was hydrogenated in the presence of 10% palladium on carbon
(50 mg) at 50 psi for 2.5 h. Purification by medium pressure
chromatography (9:1 hexanes:EtOAc to 1:1 hexanes:EtOAc to EtOAc)
provided
7-{2S-[3R-hydroxy-4-(3-trifluoromethoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-
-yl}-heptanoic acid ethyl ester (49 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 7.33 (m, 1H), 7.11 (m, 3H), 4.09 (q, 2H), 3.84 (m, 1H),
3.59 (m, 2H), 2.85 (m, 2H), 2.72 (m, 1H), 2.42-2.24 (m, 4H), 2.10
(m, 1H), 1.79 (m, 1H), 1.68-1.21 (m, 16H); MS 474.2 (M+1).
Step D:
7-{2S-[3R-Hydroxy-4-(3-trifluoromethoxy-phenyl)-butyl]-5-oxo-pyrr-
olidin-1-yl}-heptanoic acid. Analogous to the procedure described
for Compound 2A, Step E,
7-{2S-[3R-hydroxy-4-(3-trifluoromethoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-
-yl}-heptanoic acid ethyl ester (45 mg, 0.095 mmol) was hydrolyzed
with 1M NaOH (0.95 mL) in MeOH (20 mL) under reflux over 4 h to
provide the title compound of Example 2G (35 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.33 (m, 1H), 7.10 (m, 3H), 3.86 (m, 1H), 3.58
(m, 2H), 2.90 (m, 1H), 2.81 (m, 1H), 2.73 (m, 1H), 2.34 (m, 4H),
2.10 (m, 1H), 1.80 (m, 1H), 1.66-1.24 (m, 13H); MS 446.3 (M+1),
444.2 (M-1).
Compound 2H
7-{2S-[4-(3-Cyano-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-heptano-
ic acid
Step A:
7-{2R-[4-(3-Bromo-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl-
}-heptanoic acid ethyl ester. Analogous to the procedure described
for Compound 2A, Step B, the anion derived from
[3-(3-bromo-phenyl)-2-oxo-propyl]-phosphonic acid dimethyl ester
(2.90 g, 9.03 mmol) and NaH (60% in oil, 489 mg, 12.23 mmol) was
reacted with 7-(2R-formyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid
ethyl ester (assumed 11.06 mmol) over 24 h. Flash chromatography
(EtOAc to 5% MeOH in EtOAc) provided
7-{2R-[4-(3-bromo-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-y-
l}-heptanoic acid ethyl ester (2.63 g). .sup.1H NMR (CDCl.sub.3)
.delta. 7.40 (d, 1H), 7.35 (s, 1H), 7.20 (m, 1H), 7.12 (d, 1H),
6.66 (dd, 1H), 6.21 (d, 1H), 4.17 (m, 1H), 4.11 (q, 2H), 3.81 (s,
2H), 3.54 (m, 1H), 2.71 (m, 1H), 2.48-2.21 (m, 5H), 1.79 (m, 1H),
1.58 (m, 2H), 1.47-1.20 (m, 9H); MS 466.1 (M+1).
Step B:
7-{2R-[4-(3-Bromo-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrolidin-
-1-yl}-heptanoic acid ethyl ester. To a solution of
7-{2R-[4-(3-bromo-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-heptan-
oic acid ethyl ester (2.63 g, 5.66 mmol) and
(R)-2-methyl-CBS-oxazaborolidine (1M in toluene, 0.85 mL, 0.85
mmol) in CH.sub.2Cl.sub.2 (225 mL) at -45.degree. C. was added
catecholborane (1M in THF, 17.0 mL, 17.0 mmol) dropwise. The
reaction mixture was stirred at -45.degree. C. for 17 h. Aqueous
HCl (1N, 17 mL) was added and the reaction mixture was warmed to
room temperature. The organic solution was washed consecutively
with 1N HCl (1.times.100 mL), water (2.times.100 mL) and brine
(1.times.100 mL). The organic solution was dried (MgSO.sub.4),
filtered and concentrated. Purification by flash chromatography
(EtOAc to 5% MeOH in EtOAc) provided
7-{2R-[4-(3-bromo-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-h-
eptanoic acid ethyl ester (705 mg) as an approximate 95:5 ratio of
3S:3R alcohol diastereomers by .sup.1H NMR. .sup.1H NMR
(CDCl.sub.3) .delta. 7.36 (m, 2H), 7.15 (m, 2H), 5.70 (dd, 1H),
5.48 (dd, 1H), 4.40 (m, 1H), 4.10 (q, 2H), 4.03 (m, 1H), 3.46 (m,
1H), 2.81 (d, 2H), 2.72 (m, 1H), 2.39 (m, 2H), 2.27 (t, 2H), 2.20
(m, 1H), 1.84-1.22 (m, 13H).
Step C:
7-{2R-[4-(3-Cyano-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrolidin-
-1-yl}-heptanoic acid ethyl ester. Nitrogen was bubbled into a
solution of
7-{2R-[4-(3-bromo-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-h-
eptanoic acid ethyl ester (700 mg, 1.50 mmol) in DMF (2.6 mL) for 5
minutes. Zinc cyanide (108 mg, 0.92 mmol) and
tetrakis(triphenylphosphine)palladium(0) (58 mg, 0.05 mmol) were
added and nitrogen was bubbled into the reaction mixture for 5
minutes. The reaction mixture was heated at 105.degree. C. for 24
h. Additional tetrakis(triphenylphosphine)palladium(0) (58 mg,
0.050 mmol) was added and heating was continued for 1.5 h. The
reaction mixture was poured into water (50 mL) and the aqueous
solution was washed with Et.sub.2O (3.times.50 mL). The combined
ethereal layers were dried (MgSO.sub.4), filtered and concentrated
in vacuo. Medium pressure chromatography (EtOAc to 5% MeOH in EtOAc
to 10% MeOH in EtOAc) provided
7-{2R-[4-(3-cyano-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-h-
eptanoic acid ethyl ester (323 mg). .sup.1H NMR (CDCl.sub.3)
.delta.7.53 (m, 2H), 7.48-7.39 (m, 2H), 5.72 (dd, 1H), 5.51 (dd,
1H), 4.41 (m, 1H), 4.10 (q, 2H), 4.03 (m, 1H), 3.46 (m, 1H), 2.86
(m, 2H), 2.73 (m, 1H), 2.36 (m, 2H), 2.27 (t, 2H), 2.20 (m, 1H),
1.71-1.22 (m, 13H); MS 413.3 (M+1).
Step D:
7-{2S-[4-(3-Cyano-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl-
}-heptanoic acid ethyl ester. Analogous to the procedure described
for Compound 2A, Step D, a solution of
7-{2R-[4-(3-cyano-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-h-
eptanoic acid ethyl ester (150 mg, 0.36 mmol) in EtOH (13 mL) was
hydrogenated in the presence of 10% palladium on carbon (16 mg) at
45 psi for 3.5 h to provide
7-{2S-[4-(3-cyano-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-heptan-
oic acid ethyl ester (150 mg). .sup.1H NMR (CDCl.sub.3) .delta.
7.54 (m, 2H), 7.44 (m, 2H), 4.09 (q, 2H), 3.84 (m, 1H), 3.60 (m,
2H), 2.95-2.71 (m, 3H), 2.36 (m, 2H), 2.27 (t, 2H), 2.11 (m, 1H),
1.79 (m, 1H), 1.68-1.20 (m, 16H); MS 415.2 (M+1).
Step E:
7-{2S-[4-(3-Cyano-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl-
}-heptanoic acid. Analogous to the procedure described for Compound
2A, Step E,
7-{2S-[4-(3-cyano-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl-
}-heptanoic acid ethyl ester (150 mg, 0.36 mmol) was hydrolyzed
with 5M NaOH (3 mL) in EtOH (5 mL) at room temperature over 24 h to
provide the title compound of Example 2H (119 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.52 (m, 2H), 7.43 (m, 2H), 3.84 (m, 1H), 3.56
(m, 2H), 2.93-2.70 (m, 3H), 2.32 (m, 4H), 2.09 (m, 1H), 1.78 (m,
1H), 1.65-1.21 (m, 13H); MS 387.2 (M.times.1).
Compound 2I
7-(2S-{3R-Hydroxy-4-[3-(2-methoxy-ethyl)-phenyl]-butyl}-5-oxo-pyrrolidin-1-
-yl)-heptanoic acid
Step A:
7-(2R-{4-[3-(2-Methoxy-ethyl)-phenyl]-3-oxo-but-1-enyl}-5-oxo-pyr-
rolidin-1-yl)-heptanoic acid ethyl ester. Analogous to the
procedure described for Compound 2A, Step B, the anion derived from
{3-[3-(2-methoxy-ethyl)-phenyl]-2-oxo-propyl}-phosphonic acid
diethyl ester (130 mg, 0.396 mmol) and NaH (60% in oil, 17 mg,
0.425 mmol) was reacted with
7-(2R-formyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid ethyl ester
(assumed 0.461 mmol) over 24 h. Medium pressure chromatography (50%
EtOAc in hexanes to EtOAc) provided
7-(2R-{4-[3-(2-methoxy-ethyl)-phenyl]-3-oxo-but-1-enyl}-5-oxo-pyrrolidin--
1-yl)-heptanoic acid ethyl ester (101 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 7.23 (m, 1H), 7.11 (m, 1H), 7.02 (m, 2H), 6.62 (dd, 1H),
6.20 (d, 1H), 4.12 (m, 3H), 3.80 (s, 2H), 3.56 (t, 2H), 3.51 (m,
1H), 3.32 (s, 3H), 2.84 (t, 2H), 2.68 (m, 1H), 2.37 (m, 2H), 2.24
(m, 3H), 1.75 (m, 1H), 1.56 (m, 2H), 1.42-1.17 (m, 9H); MS 444.2
(M+1).
Step B:
7-(2R-{3S-Hydroxy-4-[3-(2-methoxy-ethyl)-phenyl]-but-1-enyl}-5-ox-
o-pyrrolidin-1-yl)-heptanoic acid ethyl ester. To a solution of
7-(2R-{4-[3-(2-methoxy-ethyl)-phenyl]-3-oxo-but-1-enyl}-5-oxo-pyrrolidin--
1-yl)-heptanoic acid ethyl ester (88 mg, 0.198 mmol) and
(R)-2-methyl-CBS-oxazaborolidine (1M in toluene, 0.200 mL, 0.200
mmol) in CH.sub.2Cl.sub.2 (10 mL) at -45.degree. C. was added
catecholborane (1M in THF, 0.60 mL, 0.60 mmol) dropwise. The
reaction mixture was stirred at -45.degree. C. for 24 h. Aqueous
HCl (1N, 10 mL) was added and the reaction mixture was warmed to
room temperature and was stirred for 1.5 h. The organic solution
was washed with cold 1N NaOH (3.times.15 mL) followed by brine
(1.times.20 mL). The organic solution was dried (MgSO.sub.4),
filtered and concentrated. Purification by medium pressure
chromatography (50% EtOAc in hexanes to 75% EtOAc in hexanes to
EtOAc) provided
7-(2R-{3S-hydroxy-4-[3-(2-methoxy-ethyl)-phenyl]-but-1-enyl}-5-o-
xo-pyrrolidin-1-yl)-heptanoic acid ethyl ester (45 mg) as an
approximate 4:1 mixture of 3S:3R alcohol diasteromers by .sup.1H
NMR. .sup.1H NMR (CDCl.sub.3) .delta. 7.22 (m, 1H), 7.09 (m, 1H),
7.04 (m, 2H), 5.72 (dd, 1H), 5.49 (dd, 1H), 4.38 (m, 1H), 4.10 (q,
2H), 4.02 (m, 1H), 3.58 (t, 2H), 3.46 (m, 1H), 3.34 (s, 3H),
2.87-2.68 (m, 5H), 2.41-2.24 (m, 4H), 2.18 (m, 1H), 1.70 (m, 2H),
1.59 (m, 2H), 1.48-1.21 (m, 9H); MS 446.4 (M+1).
Step C:
7-(2S-{3R-Hydroxy-4-[3-(2-methoxy-ethyl)-phenyl]-butyl}-5-oxo-pyr-
rolidin-1-yl)-heptanoic acid ethyl ester. Analogous to the
procedure described for Compound 2A, Step D, a solution of
7-(2R-{3S-hydroxy-4-[3-(2-methoxy-ethyl)-phenyl]-but-1-enyl}-5-oxo-pyrrol-
idin-1-yl)-heptanoic acid ethyl ester (43 mg, 0.0965 mmol) in EtOH
(20 mL) was hydrogenated in the presence of 10% palladium on carbon
(20 mg) at 50 psi for 18 h. Purification by medium pressure
chromatography (50% EtOAc in hexanes to EtOAc to 10% MeOH in
CH.sub.2Cl.sub.2) provided
7-(2S-{3R-hydroxy-4-[3-(2-methoxy-ethyl)-phenyl]-butyl}-5-oxo-pyrrolidin--
1-yl)-heptanoic acid ethyl ester (16 mg). MS 448.3 (M+1).
Step D:
7-(2S-{3R-Hydroxy-4-[3-(2-methoxy-ethyl)-phenyl]-butyl}-5-oxo-pyr-
rolidin-1-yl)-heptanoic acid. Analogous to the procedure described
for Compound 2A, Step E,
7-(2S-{3R-hydroxy-4-[3-(2-methoxy-ethyl)-phenyl]-butyl}-5-oxo-pyrrolidin--
1-yl)-heptanoic acid ethyl ester (15 mg, 0.034 mmol) was hydrolyzed
with 6M NaOH (0.20 mL) in EtOH (0.50 mL) at room temperature over
18 h to provide the title compound, Compound 2I (14 mg). .sup.1H
NMR (CDCl.sub.3) .delta. 7.22 (m, 1H), 7.05 (m, 3H), 3.82 (m, 1H),
3.56 (m, 4H), 3.32 (s, 3H), 2.93-2.82 (m, 3H), 2.76 (m, 1H), 2.62
(m, 1H), 2.42-2.25 (m,
Compound 2J
7-{2R-[3-Hydroxy-4-(3-phenoxy-phenyl)-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-h-
eptanoic acid
Step A:
7-{2-Oxo-5R-[3-oxo-4-(3-phenoxy-phenyl)-but-1-enyl]-pyrrolidin-1--
yl}-heptanoic acid ethyl ester. Analogous to the procedure
described for Compound 2A, Step B, the anion derived from
[2-oxo-3-(3-phenoxy-phenyl)-propyl]-phosphonic acid dimethyl ester
(633 mg, 1.98 mmol) and NaH (60% in oil, 70 mg, 1.74 mmol) was
reacted with 7-(2R-formyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid
ethyl ester (assumed 1.58 mmol) over 24 h. Medium pressure
chromatography (EtOAc) provided
7-{2-oxo-5R-[3-oxo-4-(3-phenoxy-phenyl)-but-1-enyl]-pyrrolidin-1-yl}-hept-
anoic acid ethyl ester (215 mg). .sup.1H NMR (CDCl.sub.3) .delta.
7.28 (m, 3H), 7.08 (m, 1H), 6.97 (m, 2H), 6.89 (m, 2H), 6.83 (m,
1H), 6.62 (dd, 1H), 6.19 (d, 1H), 4.13 (m, 1H), 4.08 (q, 2H), 3.79
(s, 2H), 3.51 (m, 1H), 2.68 (m, 1H), 2.35 (m, 2H), 2.24 (m, 3H),
2.24 (m, 3H), 1.75 (m, 1H), 1.54 (m, 2H), 1.43-1.20 (m, 9H).
Step B:
7-{2R-[3-hydroxy-4-(3-phenoxy-phenyl)-but-1-enyl]-5-oxo-pyrrolidi-
n-1-yl}-heptanoic acid ethyl ester. Analogous to the procedure
described for Compound 2B, Step C,
7-{2-oxo-5R-[3-oxo-4-(3-phenoxy-phenyl)-but-1-enyl]-pyrrolidin-1-yl}-hept-
anoic acid ethyl ester (215 mg, 0.451 mmol) was reacted with
NaBH.sub.4 (17 mg, 0.45 mmol) in EtOH (3 mL) at 0.degree. C. over 4
h. Purification by medium pressure chromatography (EtOAc) provided
7-{2R-[3-hydroxy-4-(3-phenoxy-phenyl)-but-1-enyl]-5-oxo-pyrrolidin-1-yl}--
heptanoic acid ethyl ester (167 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 7.33 (m, 2H), 7.25 (m, 1H), 7.10 (m, 1H), 6.99 (m, 2H),
6.93 (m, 1H), 6.86 (m, 2H), 5.72 (m, 1H), 5.45 (m, 1H), 4.37 (m,
1H), 4.10 (q, 2H), 3.47 (m, 1H), 2.82 (m, 3H), 2.35 (m, 2H), 2.26
(t, 2H), 2.15 (m, 1H), 1.70-1.21 (m, 13H).
Step C:
7-{2R-[3-Hydroxy-4-(3-phenoxy-phenyl)-but-1-enyl]-5-oxo-pyrrolidi-
n-1-yl}-heptanoic acid. Analogous to the procedure described for
Compound 2A, Step E,
7-{2R-[3-hydroxy-4-(3-phenoxy-phenyl)-but-1-enyl]-5-oxo-pyrrolidin-1-yl}--
heptanoic acid ethyl ester (29 mg, 0.060 mmol) was hydrolyzed with
2M NaOH in EtOH (4.0 mL) at room temperature over 24 h to provide
the title compound of Example 2J (20 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 7.33-7.21 (m, 3H), 7.08 (m, 1H), 6.98-6.84 (m, 5H), 5.70
(m, 1H), 5.44 (m, 1H), 4.36 (m, 1H), 4.00 (m, 1H), 3.44 (m, 1H),
2.85-2.51 (m, 3H), 2.32 (m, 4H), 2.14 (m, 1H), 1.68-1.18 (m,
10H).
Compound 2K
7-{2S-[3-Hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl}-heptan-
oic acid
Step A:
7-{2S-[3-Hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-y-
l}-heptanoic acid ethyl ester. Analogous to the procedure described
for Compound 2A, Step D, a mixture of
7-{2R-[3-hydroxy-4-(3-phenoxy-phenyl)-but-1-enyl]-5-oxo-pyrrolidin-1-yl}--
heptanoic acid ethyl ester (139 mg, 0.290 mmol), MeOH (30 mL), and
10% palladium on carbon (14 mg) was hydrogenated on a Parr shaker
at 50 psi for 18 h. Purification by medium pressure chromatography
(1:1 hexanes:EtOAc) provided
7-{2S-[3-hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl}-hepta-
noic acid ethyl ester (86 mg). .sup.1H NMR (CDCl.sub.3) .delta.
7.35-7.24 (m, 3H), 7.10 (m, 1H), 6.99 (m, 2H), 6.93 (m, 1H), 6.87
(m, 2H), 4.09 (q, 2H), 3.80 (m, 1H), 2H), 2.82 (m, 2H), 2.64 (m,
1H), 2.42-2.24 (m, 4H), 2.10 (m, 1H), 1.77 (m, 1H), 1.66-1.21 (m,
16H).
Step B:
7-{2S-[3-Hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-y-
l}-heptanoic acid. Analogous to the procedure described for
Compound 2A, Step E,
7-{2S-[3-hydroxy-4-(3-phenoxy-phenyl)-butyl]-5-oxo-pyrrolidin-1-y-
l}-heptanoic acid ethyl ester (86 mg, 1.79 mmol) was hydrolyzed
with 2N NaOH in MeOH (4 mL) over 18 h to provide the title compound
of Example 2K (62 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.33-7.23
(m, 3H), 7.09 (m, 1H), 6.98 (m, 2H), 6.91 (m, 1H), 6.86 (m, 2H),
3.80 (m, 1H), 3.56 (m, 2H), 2.88 (m, 1H), 2.77 (m, 1H), 2.64 (m,
1H), 2.38-2.28 (m, 4H), 2.09 (m, 1H), 1.77 (m, 1H), 1.64-1.21 (m,
13H).
Compound 3A
5-{3-[2S-(3-Hydroxy-4-thiophen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}--
thiophene-2-carboxylic acid
Step A:
5-{3-[2-Oxo-5R-(3-oxo-4-thiophen-2-yl-but-1-enyl)-pyrrolidin-1-yl-
]-propyl}-thioihene-2-carboxylic acid methyl ester. Analogous to
the procedure described for Compound 2A, Step B, the anion derived
from (2-oxo-3-thiophen-2-yl-propyl)-phosphonic acid dimethyl ester
(101 mg, 0.407 mmol) and NaH (60% by weight in oil, 16 mg, 0.41
mmol) was reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxyli-
c acid methyl ester (prepared from
5-[3-(2R-hydroxymethyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxy-
lic acid methyl ester analogous to the procedure described for
Compound 2A, Step A) (assumed 0.34 mmol) over 17 h. Purification by
medium pressure chromatography (1:1 hexanes:EtOAc to EtOAc)
provided
5-{3-[2-oxo-5R-(3-oxo-4-thiophen-2-yl-but-1-enyl)-pyrrolidin-1-yl]-propyl-
}-thiophene-2-carboxylic acid methyl ester (74 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.60 (d, 1H), 7.21 (m, 1H), 6.96 (m, 1H), 6.88
(m, 1H), 6.78 (d, 1H), 6.65 (dd, 1H), 6.23 (d, 1H), 4.14 (m, 1H),
4.01 (s, 2H), 3.84 (s, 3H), 3.58 (m, 1H), 2.88-2.77 (m, 3H),
2.46-2.17 (m, 3H), 1.82 (m, 3H); MS 418.0 (M+1), 416.0 (M-1).
Step B:
5-{3-[2-Oxo-5S-(3-oxo-4-thiophen-2-yl-butyl)-pyrrolidin-1-yl]-pro-
pyl}-thiophene-2-carboxylic acid methyl ester. Analogous to the
procedure described for Compound 2A, Step D,
5-{3-[2-oxo-5R-(3-oxo-4-thiophen-2-yl-but-1-enyl)-pyrrolidin-1-yl]-propyl-
}-thiophene-2-carboxylic acid methyl ester (71 mg, 0.17 mmol) was
hydrogenated in EtOH (20 mL) in the presence of 10% palladium on
carbon (50 mg) at 50 psi for 2 h. Additional catalyst was added (50
mg) and the reaction mixture was hydrogenated at 50 psi for an
additional 1 h to provide
5-{3-[2-oxo-5S-(3-oxo-4-thiophen-2-yl-butyl)-pyrrolidin-1-yl]-pro-
pyl}-thiophene-2-carboxylic acid methyl ester (63 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.61 (d, 1H), 7.22 (m, 1H), 6.97 (m, 1H), 6.88
(m, 1H), 6.80 (d, 1H), 3.88 (s, 2H), 3.84 (s, 3H), 3.65 (m, 1H),
3.52 (m, 1H), 2.95 (m, 1H), 2.81 (t, 2H), 2.48 (m, 1H), 2.30 (m,
2H), 2.07-1.80 (m, 4H), 1.55 (m, 3H); MS 419.9 (M+1), 418.0
(M-1).
Step C:
5-{3-[2S-(3-Hydroxy-4-thiophen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-
-propyl}-thiophene-2-carboxylic acid methyl ester. Analogous to the
procedure described for Compound 2B, Step C,
5-{3-[2-oxo-5S-(3-oxo-4-thiophen-2-yl-butyl)-pyrrolidin-1-yl]-propyl}-thi-
ophene-2-carboxylic acid methyl ester (60 mg, 0.143 mmol) was
reduced with NaBH.sub.4 (5 mg, 0.132 mmol) over 2 h. Purification
by preparative thin layer chromatography (EtOAc) provided
5-{3-[2S-(3-hydroxy-4-thiophen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-
-thiophene-2-carboxylic acid methyl ester (10 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.61 (d, 1H), 7.18 (d, 1H), 6.96 (m, 1H), 6.85
(d, 1H), 6.81 (d, 1H), 3.83 (s, 3H), 3.80 (m, 1H), 3.61 (m, 2H),
3.00 (m, 2H), 2.89 (m, 1H), 2.83 (t, 2H), 2.34 (m, 2H), 2.10 (m,
1H), 1.98-1.23 (m, 8H); MS 422.2 (M+1).
Step D:
5-{3-[2S-(3-Hydroxy-4-thiophen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-
-propyl}-thiophene-2-carboxylic acid. Analogous to the procedure
described for Compound 2A, Step E,
5-{3-[2S-(3-hydroxy-4-thiophen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-
-thiophene-2-carboxylic acid methyl ester (10 mg, 0.024 mmol) was
hydrolyzed with NaOH (1M, 0.03 mL) in MeOH (5 mL) over 29 h to
provide the title compound, Compound 3A (10 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.68 (d, 1H), 7.18 (m, 1H), 6.96 (m, 1H), 6.85
(m, 2H), 3.80 (m, 1H), 3.63 (m, 2H), 3.01 (m, 2H), 2.91 (m, 1H),
2.85 (t, 2H), 2.36 (m, 2H), 2.11 (m, 1H), 2.00-1.18 (m, 8H).
Compound 3B
5-(3-{2S-[4-(4-Chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-thiophene-2-carboxylic acid
Step A:
5-(3-{2R-[4-(4-Chloro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous
to the procedure described for Compound 2A, Step B, the anion
derived from [3-(4-chloro-phenyl)-2-oxo-propyl]-phosphonic acid
dimethyl ester (113 mg, 0.407 mmol) and NaH (60% by weight in oil,
16 mg, 0.41 mmol) was reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxylic
acid methyl ester (assumed 0.34 mmol) over 17 h. Purification by
medium pressure chromatography (1:1 hexanes:EtOAc to EtOAc)
provided
5-(3-{2R-[4-(4-chloro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid methyl ester (94 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.61 (d, 1H), 7.29 (m, 2H), 7.10 (d, 2H), 6.78
(d, 1H), 6.62 (dd, 1H), 6.18 (d, 1H), 4.13 (m, 1H), 3.84 (s, 3H),
3.79 (s, 2H), 3.56 (m, 1H), 2.87-2.77 (m, 3H), 2.47-2.16 (m, 3H),
1.80 (m, 3H).
Step B:
5-(3-{2S-[4-(4-Chloro-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}-
-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous to the
procedure described for Compound 2A, Step D,
5-(3-{2R-[4-(4-chloro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid methyl ester (91 mg, 0.204 mmol)
was hydrogenated in EtOH (20 mL) in the presence of 10% palladium
on carbon (50 mg) at 50 psi for 2 h to provide
5-(3-{2S-[4-(4-chloro-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}-propyl)-
-thiophene-2-carboxylic acid methyl ester (84 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.61 (d, 1H), 7.30 (d, 2H), 7.11 (d, 2H), 6.80
(d, 1H), 3.84 (s, 3H), 3.66 (s, 2H), 3.64 (m, 1H), 3.51 (m, 1H),
2.94 (m, 1H), 2.81 (t, 2H), 2.42 (m, 2H), 2.29 (m, 2H), 2.04-1.79
(m, 4H), 1.56 (m, 2H); MS 448.0 (M+1), 446.0 (M-1).
Step C:
5-(3-{2S-[4-(4-Chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous to
the procedure described for Compound 2B, Step C,
5-(3-{2S-[4-(4-chloro-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}-propyl)-
-thiophene-2-carboxylic acid methyl ester (81 mg, 0.181 mmol) was
reduced with NaBH.sub.4 (7 mg, 0.181 mmol) over 2 h. Purification
by preparative thin layer chromatography (EtOAc, 2.times.) provided
5-(3-{2S-[4-(4-chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid methyl ester (54 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.61 (d, 1H), 7.28 (d, 2H), 7.12 (d, 2H), 6.81
(d, 1H), 3.82 (s, 3H), 3.77 (m, 1H), 3.60 (m, 2H), 2.99 (m, 1H),
2.83 (t, 2H), 2.77 (m, 1H), 2.62 (m, 1H), 2.34 (m, 2H), 2.09 (m,
1H), 1.97-1.30 (m, 8H); MS 450.0 (M+1).
Step D:
5-(3-{2S-[4-(4-Chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-thiophene-2-carboxylic acid. Analogous to the
procedure described for Compound 2A, Step E,
5-(3-{2S-[4-(4-chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid methyl ester (52 mg, 0.116 mmol)
was hydrolyzed with NaOH (1M, 0.14 mL) in MeOH (5 mL) under reflux
over 29 h to provide
5-(3-{2S-[4-(4-chloro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid (16 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 7.67 (d, 1H), 7.28 (d, 2H), 7.12 (d, 2H), 6.84 (d, 1H),
3.78 (m, 1H), 3.62 (m, 1H), 3.01 (m, 1H), 2.85 (t, 2H), 2.77 (m,
1H), 2.63 (m, 1H), 2.36 (m, 2H), 2.10 (m, 1H), 1.90 (m, 3H), 1.75
(m, 1H), 1.69-1.24 (m, 4H); MS 434.0 (M-1).
Compound 3C
5-(3-{2S-[3-Hydroxy-4-(2-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-thiophene-2-carboxylic acid
Step A:
5-(3-{2-Oxo-5R-[3-oxo-4-(2-trifluoromethyl-phenyl)-but-1-enyl]-py-
rrolidin-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester.
Analogous to the procedure described for Compound 2A, Step B, the
anion derived from
[2-oxo-3-(2-trifluoromethyl-phenyl)-propyl]-phosphonic acid
dimethyl ester (74 mg, 0.239 mmol) and NaH (60% by weight in oil,
10 mg, 0.239 mmol) was reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxylic
acid methyl ester (assumed 0.239 mmol) over 17 h. Purification by
medium pressure chromatography (1:1 hexanes:EtOAc to EtOAc)
provided
5-(3-{2-oxo-5R-[3-oxo-4-(2-trifluoromethyl-phenyl)-but-1-enyl]-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (32 mg).
.sup.1H NMR (CDCl.sub.3) .delta. 7.66 (d, 1H), 7.60 (m, 1H), 7.51
(m, 1H), 7.39 (m, 1H), 7.28 (m, 1H), 6.79 (m, 1H), 6.64 (dd, 1H),
6.22 (d, 1H), 4.16 (m, 1H), 3.83 (s, 3H), 3.78 (s, 2H), 3.60 (m,
1H), 2.93-2.79 (m, 3H), 2.48-2.20 (m, 3H), 1.83 (m, 3H); MS 479.9
(M+1). 478.0 (M-1).
Step B:
5-(3-{2-Oxo-5S-[3-oxo-4-(2-trifluoromethyl-phenyl)-butyl]-pyrroli-
din-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester.
Analogous to the procedure described for Compound 2A, Step D,
5-(3-{2-oxo-5R-[3-oxo-4-(2-trifluoromethyl-phenyl)-but-1-enyl]-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (29 mg,
0.060 mmol) was hydrogenated in EtOH (20 mL) in the presence of 10%
palladium on carbon (40 mg) at 50 psi for 2 h to provide
5-(3-{2-oxo-5S-[3-oxo-4-(2-trifluoromethyl-phenyl)-butyl]-pyrrolidin-1-yl-
}-propyl)-thiophene-2-carboxylic acid methyl ester (29 mg). .sup.1H
NMR (CDCl.sub.3) .delta. 7.66 (d, 1H), 7.59 (m, 1H), 7.52 (m, 1H),
7.39 (m, 1H), 7.27 (m, 1H), 6.80 (d, 1H), 3.83 (s, 3H), 3.78 (s,
2H), 3.64 (m, 1H), 3.55 (m, 1H), 2.97 (m, 1H), 2.81 (t, 2H), 2.48
(m, 1H), 2.33 (m, 2H), 2.05 (m, 2H), 1.87 (m, 2H), 1.56 (m, 3H); MS
482.0 (M+1), 480.0 (M-1).
Step C:
5-(3-{2S-[3-Hydroxy-4-(2-trifluoromethyl-phenyl)-butyl]-5-oxo-pyr-
rolidin-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester.
Analogous to the procedure described for Compound 2B, Step C,
5-(3-{2-oxo-5S-[3-oxo-4-(2-trifluoromethyl-phenyl)-butyl]-pyrrolidin-1-yl-
}-propyl)-thiophene-2-carboxylic acid methyl ester (26 mg, 0.054
mmol) was reduced with NaBH.sub.4 (2 mg, 0.054 mmol) over 2 h.
Purification by preparative thin layer chromatography (EtOAc)
provided
5-(3-{2S-[3-hydroxy-4-(2-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (10 mg).
.sup.1H NMR (CDCl.sub.3) .delta. 7.65 (d, 1H), 7.59 (m, 1H), 7.49
(m, 1H), 7.36 (m, 2H), 6.81 (d, 1H), 3.81 (s, 3H), 3.81 (m, 1H),
3.62 (m, 2H), 3.02 (m, 2H), 2.83 (t, 2H), 2.78 (m, 1H), 2.34 (m,
2H), 2.12 (m, 1H), 2.01-1.35 (m, 8H); MS 484.0 (M+1).
Step D:
5-(3-{2S-[3-Hydroxy-4-(2-trifluoromethyl-phenyl)-butyl]-5-oxo-pyr-
rolidin-1}-propyl)-thiophene-2-carboxylic acid. Analogous to the
procedure described for Compound 2A, Step E,
5-(3-{2S-[3-hydroxy-4-(2-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (10 mg,
0.0207 mmol) was hydrolyzed with NaOH (1M, 0.07 mL) in MeOH (5 mL)
heated under reflux for 29 h to provide
5-(3-{2S-[3-hydroxy-4-(2-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid (13 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.66 (m, 1H), 7.50 (m, 1H), 7.37 (m, 3H), 6.84
(d, 1H), 3.83 (m, 1H), 3.64 (m, 2H), 3.04 (m, 2H), 2.85 (t, 2H),
2.78 (m, 1H), 2.37 (m, 2H), 2.12 (m, 1H), 2.02-1.24 (m, 8H); MS
470.1 (M+1), 468.0 (M-1).
Compound 3D
5-(3-{2S-[4-(4-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-thiophene-2-carboxylic acid
Step A:
5-(3-{2R-[4-(4-Fluoro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous
to the procedure described for Compound 2A, Step B, the anion
derived from [3-(4-fluoro-phenyl)-2-oxo-propyl]-phosphonic acid
dimethyl ester (106 mg, 0.407 mmol) and NaH (60% by weight in oil,
16 mg, 0.407 mmol) was reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxylic
acid methyl ester (assumed 0.407 mmol) over 17 h. Purification by
medium pressure chromatography (1:1 hexanes:EtOAc to EtOAc)
provided
5-(3-{2R-[4-(4-fluoro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid methyl ester (77 mg). .sup.1H NMR
(CDCl.sub.3) .delta. (d, 1H), 7.16 (m, 2H), 7.00 (m, 2H), 6.77 (d,
1H), 6.62 (dd, 1H), 6.19 (d, 1H), 4.13 (m, 1H), 3.84 (s, 3H), 3.79
(s, 2H), 3.57 (m, 1H), 2.87-2.77 (m, 3H), 2.37 (m, 2H), 2.20 (m,
1H), 1.80 (m, 3H); MS 430.0 (M+1), 428.1 (M-1).
Step B:
5-(3-{2S-[4-(4-Fluoro-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}-
-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous to the
procedure described for Compound 2A, Step D,
5-(3-{2R-[4-(4-fluoro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid methyl ester (74 mg, 0.172 mmol)
was hydrogenated in EtOH (20 mL) in the presence of 10% palladium
on carbon (50 mg) at 50 psi for 2 h to provide
5-(3-{2S-[4-(4-fluoro-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}-propyl)-
-thiophene-2-carboxylic acid methyl ester (72 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.61 (d, 1H), 7.14 (m, 2H), 7.01 (m, 2H), 6.80
(d, 1H), 3.84 (s, 3H), 3.66 (s, 2H), 3.64 (m, 1H), 3.51 (m, 1H),
2.94 (m, 1H), 2.81 (t, 2H), 2.43 (m, 2H), 2.30 (m, 2H), 2.05-1.79
(m, 4H), 1.56 (m, 2H); MS 432.0 (M+1), 430.1 (M-1).
Step C:
5-(3-{2S-[4-(4-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous to
the procedure described for Compound 2B, Step C,
5-(3-{2S-[4-(4-fluoro-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}-propyl)-
-thiophene-2-carboxylic acid methyl ester (69 mg, 0.160 mmol) was
reduced with NaBH.sub.4 (6 mg, 0.160 mmol) over 2 h. Purification
by preparative thin layer chromatography (EtOAc) provided
5-(3-{2S-[4-(4-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid methyl ester (37 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.61 (d, 1H), 7.15 (m, 2H), 7.00 (m, 2H), 6.81
(d, 1H), 3.82 (s, 3H), 3.75 (m, 1H), 3.60 (m, 2H), 2.99 (m, 1H),
2.83 (t, 2H), 2.77 (m, 1H), 2.34 (m, 2H), 2.10 (m, 1H), 2.00-1.80
(m, 4H), 1.75 (m, 1H), 1.68-1.34 (m, 4H); MS 434.3 (M+1).
Step D:
5-(3-{2S-[4-(4-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-thiophene-2-carboxylic acid. Analogous to the
procedure described for Compound 2A, Step E,
5-(3-{2S-[4-(4-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid methyl ester (35 mg, 0.0807 mmol)
was hydrolyzed with NaOH (1M, 0.10 mL) in MeOH (5 mL) heated under
reflux over 29 h to provide
5-(3-{2S-[4-(4-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid (36 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 7.67 (d, 1H), 7.15 (m, 2H), 7.00 (m, 2H), 6.84 (d, 1H),
3.77 (m, 1H), 3.62 (m, 2H), 3.01 (m, 1H), 2.85 (t, 2H), 2.78 (m,
1H), 2.62 (m, 1H), 2.36 (m, 2H), 2.10 (m, 1H), 2.00-1.72 (m, 4H),
1.69-1.34 (m, 4H); MS 420.1 (M+1), 417.7 (M-1).
Compound 3E
5-(3-{2S-[4-(4-Fluoro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid
Step A:
5-(3-{2R-[4-(4-Fluoro-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrol-
idin-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. To a
solution of
5-(3-{2R-[4-(4-fluoro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-
-propyl)-thiophene-2-carboxylic acid methyl ester (20 mg, 0.047
mmol) and (R)-2-methyl-CBS-oxazaborolidine (1M in toluene, 0.047
mL, 0.047 mmol) in anhydrous toluene (3.0 mL) at -45.degree. C. was
added catecholborane (1M in THF, 0.14 mL, 0.14 mmol) dropwise. The
reaction mixture was stirred at -45.degree. C. for 17 h. Methanol
(1 mL) was added and the reaction mixture was warmed to room
temperature and was concentrated in vacuo. The residue was
dissolved in CHCl.sub.3 and the organic solution was washed with 1M
NaOH (4.times.5 mL), 1M HCl (1.times.5 mL), and water (1.times.5
mL). The organic solution was dried (MgSO.sub.4), filtered and
concentrated. Purification by preparative thin layer chromatography
(EtOAc) provided
5-(3-{2R-[4-(4-fluoro-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrolidin-1-y-
l}-propyl)-thiophene-2-carboxylic acid methyl ester as an
approximate 39:1 ratio of 3S:3R alcohol diastereomers by HPLC. MS
432.1 (M+1).
Step B:
5-(3-{2S-[4-(4-Fluoro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous
to the procedure described for Compound 2A, Step D,
5-(3-{2R-[4-(4-fluoro-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrolidin-1-y-
l}-propyl)-thiophene-2-carboxylic acid methyl ester (15 mg, 0.035
mmol) was hydrogenated in ethanol (10 mL) in the presence of 10%
palladium on carbon (5 mg) at 50 psi for 2 h to provide
5-(3-{2S-[4-(4-fluoro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid methyl ester (11 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.60 (d, 1H), 7.14 (m, 2H), 7.00 (m, 2H), 6.81
(d, 1H), 3.82 (s, 3H), 3.77 (m, 1H), 3.60 (m, 2H), 3.00 (m, 1H),
2.83 (t, 2H), 2.76 (dd, 1H), 2.63 (dd, 1H), 2.34 (m, 2H), 2.08 (m,
1H), 1.98-1.42 (m, 8H); MS 434.1 (M+1).
Step C:
5-(3-{2S-[4-(4-Fluoro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid. Analogous to the
procedure described for Compound 2A, Step E,
5-(3-{2S-[4-(4-fluoro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid methyl ester (11 mg, 0.0254 mmol)
was hydrolyzed with NaOH (1M, 0.25 mL) in MeOH (4 mL) heated under
reflux for 3 h to provide
5-(3-{2S-[4-(4-fluoro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid (9 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 7.67 (d, 1H), 7.14 (m, 2H), 6.99 (m, 2H), 6.83 (d, 1H),
3.78 (m, 1H), 3.62 (m, 2H), 3.02 (m, 1H), 2.85 (t, 2H), 2.76 (dd,
1H), 2.64 (dd, 1H), 2.37 (m, 2H), 2.09 (m, 1H), 2.00-1.42 (m, 8H);
MS 420.1 (M+1), 418.0 (M-1).
Compound 3F
5-{3-[2S-(3-Hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl-
}-thiophene-2-carboxylic acid
Step A:
5-{3-[2R-(4-Naphthalen-2-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1--
yl]-propyl}-thiophene-2-carboxylic acid tert-butyl ester. Analogous
to the procedure described for Compound 2A, Step B, the anion
derived from (3-naphthalen-2-yl-2-oxo-propyl)-phosphonic acid
dimethyl ester (208 mg, 0.71 mmol) and NaH (60% by weight in oil,
26 mg, 0.65 mmol) was reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxyli-
c acid tert-butyl ester (assumed 0.589 mmol) over 18 h.
Purification by medium pressure chromatography (1:1 hexanes:EtOAc
to EtOAc) provided
5-{3-[2R-(4-naphthalen-2-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-prop-
yl}-thiophene-2-carboxylic acid tert-butyl ester (181 mg). .sup.1H
NMR (CDCl.sub.3) .delta. 7.79 (m, 3H), 7.65 (s, 1H), 7.47 (m, 3H),
7.29 (m, 1H), 6.63 (m, 2H), 6.22 (d, 1H), 4.08 (m, 1H), 3.98 (s,
2H), 3.49 (m, 1H), 2.73 (m, 1H), 2.63 (m, 2H), 2.36 (m, 2H), 2.19
(m, 1H), 1.72 (m, 3H), 1.54 (s, 9H); MS 504.1 (M+1), 502.0
(M-1).
Step B:
5-{3-[2S-(4-Naphthalen-2-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-P-
ropyl}-thiophene-2-carboxylic acid tert-butyl ester. Analogous to
the procedure described for Compound 2A, Step D,
5-{3-[2R-(4-naphthalen-2-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-prop-
yl}-thiophene-2-carboxylic acid tert-butyl ester (178 mg, 0.353
mmol) was hydrogenated in EtOH (40 mL) in the presence of 10%
palladium on carbon (75 mg) at 50 psi for 3 h. Purification by
medium pressure chromatography (1:1 hexanes:EtOAc to EtOAc)
provided
5-{3-[2S-(4-naphthalen-2-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-t-
hiophene-2-carboxylic acid tert-butyl ester (144 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.80 (m, 3H), 7.66 (s, 1H), 7.48 (m, 3H), 7.30
(m, 1H), 6.74 (d, 1H), 3.85 (s, 2H), 3.59 (m, 1H), 3.48 (m, 1H),
2.89 (m, 1H), 2.73 (t, 2H), 2.47 (m, 2H), 2.26 (m, 2H), 2.04-1.74
(m, 4H), 1.53 (s, 9H), 1.50 (m, 2H); MS 506.1 (M+1), 503.8
(M-1).
Step C:
5-{3-[2S-(3-Hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-y-
l]-propyl}-thiophene-2-carboxylic acid tert-butyl ester. Analogous
to the procedure described for Compound 2B, Step C,
5-{3-[2S-(4-naphthalen-2-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-t-
hiophene-2-carboxylic acid tert-butyl ester (142 mg, 0.281 mmol)
was reduced with NaBH.sub.4 (11 mg, 0.281 mmol) over 2 h.
Purification by medium pressure chromatography (1:1 hexanes:EtOAc
to EtOAc) provided
5-{3-[2S-(3-hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-propy-
l}-thiophene-2-carboxylic acid tert-butyl ester (125 mg). .sup.1H
NMR (CDCl.sub.3) .delta. 7.79 (m, 3H), 7.65 (s, 1H), 7.52 (d, 1H),
7.46 (m, 2H), 7.32 (d, 1H), 6.76 (d, 1H), 3.90 (m, 1H), 3.62 (m,
2H), 2.98 (m, 2H), 2.81 (m, 3H), 2.34 (m, 2H), 2.10 (m, 1H),
2.04-1.75 (m, 2H), 1.70-1.36 (m, 6H), 1.52 (s, 9H); MS
508.0(M+1).
Step D:
5-{3-[2S-(3-Hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-y-
l]-propyl}-thiophene-2-carboxylic acid. To a solution of
5-{3-[2S-(3-hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-propy-
l}-thiophene-2-carboxylic acid tert-butyl ester (123 mg, 0.242
mmol) in CH.sub.2Cl.sub.2 (20 mL) at 0.degree. C. was added TFA
(0.19 mL, 0.247 mmol). The reaction mixture was stirred at room
temperature for 23 h and was concentrated in vacuo. The residue was
purified by preparative thin layer chromatography (EtOAc) to
provide the title compound, Compound 3F (47 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.78 (m, 3H), 7.63 (m, 2H), 7.44 (m, 2H), 7.31
(m, 1H), 6.78 (m, 1H), 3.89 (m, 1H), 3.57 (m, 2H), 2.94 (m, 2H),
2.79 (m, 3H), 2.32 (m, 2H), 2.10-1.17 (m, 9H); MS 452.3 (M+1),
450.2 (M-1).
Compound 3G
5-{3-[2S-(4-Biphenyl-3-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}--
thiophene-2-carboxylic acid
Step A:
5-{3-[2R-(4-Biphenyl-3-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl-
]propyl}-thiophene-2-carboxylic acid methyl ester. Analogous to the
procedure described for Compound 2A, Step B, the anion derived from
(3-biphenyl-3-yl-2-oxo-propyl)-phosphonic acid dimethyl ester
(3.217 g, 10.09 mmol) and NaH (60% by weight in oil, 404 mg, 10.09
mmol) was reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxylic
acid methyl ester (assumed 10.09 mmol) over 17 h. Purification by
medium pressure chromatography (solvent gradient 9:1 hexanes:EtOAc
to EtOAc) provided
5-{3-[2R-(4-biphenyl-3-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-y-
l]-propyl}-thiophene-2-carboxylic acid methyl ester (4.0 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.56 (m, 3H), 7.49 (m, 1H), 7.42
(m, 4H), 7.34 (m, 1H), 7.16 (d, 1H), 6.73 (d, 1H), 6.62 (dd, 1H),
6.22 (d, 1H), 4.11 (m, 1H), 3.88 (s, 2H), 3.82 (s, 3H), 3.54 (m,
1H), 2.79 (m, 1H), 2.73 (t, 2H), 2.36 (m, 2H), 2.20 (m, 1H), 1.76
(m, 3H); MS 488.1 (M+1), 486.0 (M-1).
Step B:
5-{3-[2S-(4-Biphenyl-3-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-pro-
pyl}-thiophene-2-carboxylic acid methyl ester. Analogous to the
procedure described for Compound 2A, Step D, a mixture of
5-{3-[2R-(4-biphenyl-3-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-propyl-
}-thiophene-2-carboxylic acid methyl ester (3.535 g, 7.25 mmol),
10% palladium on carbon (750 mg), and EtOH (250 mL) was
hydrogenated at 50 psi for 2 h to provide
5-{3-[2S-(4-biphenyl-3-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-thi-
ophene-2-carboxylic acid methyl ester which was used without
further purification in Step C. MS 490.1 (M+1).
Step C:
5-{3-[2S-(4-Biphenyl-3-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]-
-propyl}-thiophene-2-carboxylic acid ethyl ester. Analogous to the
procedure described for Compound 2B, Step C,
5-{3-[2S-(4-biphenyl-3-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-thi-
ophene-2-carboxylic acid methyl ester (7.25 mmol) was treated with
NaBH.sub.4 (274 mg, 7.25 mmol) in EtOH at room temperature for 1 h.
Purification by medium pressure chromatography (1:1 hexanes:EtOAc
to EtOAc) provided
5-{3-[2S-(4-biphenyl-3-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-
-thiophene-2-carboxylic acid ethyl ester (1.68 g). .sup.1H NMR
(CDCl.sub.3) .delta. 7.58 (m, 3H), 7.40 (m, 6H), 7.17 (d, 1H), 6.79
(d, 1H), 4.27 (q, 2H), 3.85 (m, 1H), 3.62 (m, 2H), 3.00 (m, 1H),
2.86 (m, 3H), 2.71 (m, 1H), 2.34 (m, 2H), 2.10 (m, 1H), 2.01-1.75
(m, 4H), 1.70-1.35 (m, 4H), 1.31 (t, 3H); MS 506.1 (M+1).
Step D:
5-{3-[2S-(4-Biphenyl-3-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]-
-propyl}-thiophene-2-carboxylic acid. Analogous to the procedure
described for Compound 2A, Step E,
5-{3-[2S-(4-biphenyl-3-yl-3-hydroxy-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-
-thiophene-2-carboxylic acid ethyl ester (1.882 g, 3.72 mmol) was
hydrolyzed with NaOH (1M, 5.6 mL) in MeOH (100 mL) over 3 h under
reflux to provide the title compound of Example 3G (1.741 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.66 (d, 1H), 7.56 (d, 2H), 7.40
(m, 6H), 7.17 (d, 1H), 6.82 (d, 1H), 3.85 (m, 1H), 3.63 (m, 2H),
3.02 (m, 1H), 2.86 (m, 3H), 2.72 (m, 1H), 2.36 (m, 2H), 2.11 (m,
1H), 2.01-1.75 (m, 4H), 1.71-1.35 (m, 4H); MS 478.1 (M+1), 476.0
(M-1).
Compound 3H
5-(3-{2S-[4-(3-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-thiophene-2-carboxylic acid
Step A:
5-(3-{2R-[4-(3-Fluoro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous
to the procedure described for Compound 2A, Step B, the anion
derived from [3-(3-fluoro-phenyl)-2-oxo-propyl]-phosphonic acid
dimethyl ester (3.236 g, 12.4 mmol) and NaH (60% in oil, 458 mg,
11.4 mmol) was reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxylic
acid methyl ester (assumed 10.4 mmol) over 18 h. Purification by
medium pressure chromatography eluting with 20% EtOAc in hexanes to
80% EtOAc in hexanes followed by a second column eluting with 20%
acetone in toluene to 30% acetone in toluene provided
5-(3-{2R-[4-(3-fluoro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid methyl ester (2.95 g). .sup.1H
NMR (CDCl.sub.3) .delta. 7.60 (d, 1H), 7.27 (m, 1H), 6.92 (m, 3H),
6.76 (d, 1H), 6.60 (dd, 1H), 6.18 (d, 1), 4.12 (m, 1H), 3.83 (s,
3H), 3.80 (s, 2H), 3.56 (m, 1H), 2.82 (m, 1H), 2.77 (t, 2H), 2.37
(m, 2H), 2.22 (m, 1H), 1.78 (m, 3H).
Step B:
5-(3-{2S-[4-(3-Fluoro-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}-
-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous to the
procedure described for Compound 2A, Step D,
5-(3-{2R-[4-(3-fluoro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid methyl ester (2.95 g, 6.87 mmol)
was hydrogenated in MeOH (60 mL) in the presence of 10% palladium
on carbon (500 mg) at 50 psi for 2 h. Purification by medium
pressure chromatography (50% EtOAc in hexanes to EtOAc) provided
5-(3-{2S-[4-(3-fluoro-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}-propyl)-
-thiophene-2-carboxylic acid methyl ester (2.60 g). .sup.1H NMR
(CDCl.sub.3) .delta. 7.60 (d, 1H), 7.28 (m, 1H), 6.92 (m, 3H), 6.79
(d, 1H), 3.82 (s, 3H), 3.67 (s, 2H), 3.62 (m, 1H), 3.50 (m, 1H),
2.93 (m, 1H), 2.80 (t, 2H), 2.43 (m, 2H), 2.27 (m, 2H), 2.04-1.76
(m, 4H), 1.50 (m, 2H); MS 432.2 (M+1), 430.1 (M-1).
Step C:
5-(3-{2S-[4-(3-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous to
the procedure described for Compound 2B, Step C,
5-(3-{2S-[4-(3-fluoro-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}-propyl)-
-thiophene-2-carboxylic acid methyl ester (2.60 g, 6.03 mmol) was
reacted with NaBH.sub.4 (114 mg, 3.01 mmol) in MeOH (30 mL) at
0.degree. C. for 3 h. Purification by medium pressure
chromatography (EtOAc to 2% MeOH in CH.sub.2Cl.sub.2) provided
5-(3-{2S-[4-(3-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid methyl ester (2.43 g). MS 434.0
(M+1).
Step D:
5-(3-{2S-[4-(3-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-thiophene-2-carboxylic acid. Analogous to the
procedure described for Compound 2A, Step E,
5-(3-{2S-[4-(3-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid methyl ester (2.43 g) was
hydrolyzed with 2N NaOH in MeOH (30 mL) over 18 h to provide
5-(3-{2S-[4-(3-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid (2.06 g).
Step E: Sodium salt of
5-(3-{2S-[4-(3-Fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-Pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid. Analogous to the procedure
described for Compound 2D, Step E,
5-(3-{2S-[4-(3-fluoro-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid (2.058 g, 4.905 mmol) was reacted
with NaHCO.sub.3 (412 mg, 4.906 mmol) to yield the sodium salt of
the title compound of Example 3H. .sup.1H NMR (CD.sub.3OD) .delta.
7.35 (d, 1H), 7.26 (m, 1H), 6.96 (m, 3H), 6.75 (d, 1H), 3.76 (m,
1H), 3.67 (m, 1H), 3.57 (m, 1H), 3.02 (m, 1H), 2.76 (m, 3H), 2.30
(m, 2H), 2.10 (m, 1H), 1.98-1.28 (m, 9H).
Compound 3I
5-(3-{2S-[4-(4-Ethyl-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-propy-
l)-thiophene-2-carboxylic acid
Step A:
5-(3-{2R-[4-(4-Ethyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous to
the procedure described for Compound 2A, Step B, the anion derived
from [3-(4-ethyl-phenyl)-2-oxo-propyl]-phosphonic acid diethyl
ester (274 mg, 0.915 mmol) and NaH (60% by weight in oil, 41 mg,
1.01 mmol) was reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxyli-
c acid methyl ester (assumed 1.01 mmol) over 18 h. Purification by
medium pressure chromatography (1:1 hexanes:EtOAc to EtOAc)
provided
5-(3-{2R-[4-(4-ethyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-thi-
ophene-2-carboxylic acid methyl ester (227 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.59 (d, 1H), 7.13 (d, 2H), 7.07 (d, 2H), 6.75
(d, 1H), 6.58 (dd, 1H), 6.18 (d, 1H), 4.10 (m, 1H), 3.83 (s, 3H),
3.77 (s, 2H), 3.53 (m, 1H), 2.78 (m, 3H), 2.59 (q, 2H), 2.36 (m,
2H), 2.19 (m, 1H), 1.76 (m, 3H), 1.19 (t, 3H); MS 440.2 (M+1).
Step B:
5-(3-{2S-[4-(4-Ethyl-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}--
propyl)-thiophene-2-carboxylic acid methyl ester. Analogous to the
procedure described for Compound 2A, Step D,
5-(3-{2R-[4-(4-ethyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid methyl ester (227 mg, 0.517 mmol)
was hydrogenated in MeOH (30 mL) in the presence of 10% palladium
on carbon at 50 psi for 1.5 h. Purification by medium pressure
chromatography (1:1 hexanes:EtOAc to EtOAc) provided
5-(3-{2S-[4-(4-ethyl-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}-propyl)--
thiophene-2-carboxylic acid methyl ester (119 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.62 (d, 1H), 7.16 (d, 2H), 7.10 (d, 2H), 6.81
(d, 1H), 3.84 (s, 3H), 3.65 (s, 2H), 3.63 (m, 1H), 3.49 (m, 1H),
2.95 (m, 1H), 2.80 (t, 2H), 2.62 (q, 2H), 2.43 (m, 2H), 2.31 (m,
2H), 2.06-1.79 (m, 4H), 1.48 (m, 2H), 1.21 (t, 3H); MS 442.2
(M+1).
Step C:
5-(3-{2S-[4-(4-Ethyl-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1--
yl}-propyl)-thioihene-2-carboxylic acid methyl ester. Analogous to
the procedure described for Compound 2B, Step C,
5-(3-{2S-[4-(4-ethyl-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl}-propyl)--
thiophene-2-carboxylic acid methyl ester (109 mg, 0.247 mmol) was
reduced with NaBH.sub.4 (5 mg, 0.132 mmol) in MeOH (7 mL) at
0.degree. C. to room temperature over 3 h. Purification by medium
pressure chromatography (1:1 hexanes:EtOAc to EtOAc) provided
5-(3-{2S-[4-(4-ethyl-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-thiophene-2-carboxylic acid methyl ester (77 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.61 (d, 1H), 7.16 (d, 2H), 7.10 (d, 2H), 6.81
(d, 1H), 3.83 (s, 3H), 3.77 (m, 1H), 3.62 (m, 2H), 3.01 (m, 1H),
2.83 (t, 2H), 2.77 (m, 1H), 2.60 (m, 3H), 2.35 (m, 2H), 2.09 (m,
1H), 1.99-1.34 (m, 8H), 1.22 (t, 3H); MS 444.3 (M+1).
Step D:
5-(3-{2S-[4-(4-Ethyl-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1--
yl}-propyl)-thiophene-2-carboxylic acid. Analogous to the procedure
described for Compound 2A, Step E,
5-(3-{2S-[4-(4-ethyl-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-prop-
yl)-thiophene-2-carboxylic acid methyl ester (76 mg) was hydrolyzed
with 2N NaOH in MeOH (7 mL) over 18 h to provide the title compound
of Example 3(58 mg). .sup.1H NMR (CD.sub.3OD) .delta. 7.57 (m, 1H),
7.08 (d, 4H), 6.88 (d, 1H), 3.72 (m, 1H), 3.63 (m, 1H), 3.52 (m,
1H), 2.99 (m, 1H), 2.81 (t, 2H), 2.68 (m, 2H), 2.56 (q, 2H), 2.27
(m, 2H), 2.06 (m, 1H), 1.95-1.25 (m, 6H), 1.16 (t, 3H); MS 430.3
(M+1), 428.5 (M-1).
Compound 3J
5-(3-{2S-[4-(4-Fluoro-3-methyl-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin-1-
-yl}-propyl)-thiophene-2-carboxylic acid
Step A:
5-(3-{2R-[4-(4-Fluoro-3-methyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-py-
rrolidin-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester.
Analogous to the procedure described for Compound 2A, Step B, the
anion derived from
[3-(4-fluoro-3-methyl-phenyl)-2-oxo-propyl]-phosphonic acid diethyl
ester (273 mg, 0.903 mmol) and NaH (60% by weight in oil, 41 mg,
1.01 mmol) was reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxylic
acid methyl ester (assumed 1.01 mmol) over 18 h. Purification by
medium pressure chromatography (20% EtOAc in hexanes to EtOAc)
provided
5-(3-{2R-[4-(4-fluoro-3-methyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (174 mg).
.sup.1H NMR (CDCl.sub.3) .delta. 7.59 (d, 1H), 6.97 (d, 1H), 6.93
(d, 2H), 6.76 (d, 1H), 6.60 (dd, 1H), 6.18 (d, 1H), 4.11 (m, 1H),
3.82 (s, 3H), 3.73 (s, 2H), 3.56 (m, 1H), 2.82 (m, 1H), 2.77 (t,
2H), 2.36 (m, 2H), 2.22 (s, 3H), 2.19 (m, 1H), 1.78 (m, 3H); MS
444.2 (M+1); 442.2 (M-1).
Step B:
5-(3-{2S-[4-(4-Fluoro-3-methyl-phenyl)-3-oxo-butyl]-5-oxo-pyrroli-
din-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester.
Analogous to the procedure described for Compound 2A, Step D,
5-(3-{2R-[4-(4-fluoro-3-methyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (174 mg,
0.392 mmol) was hydrogenated in MeOH (30 mL) in the presence of 10%
palladium on carbon (70 mg) at 50 psi for 1.5 h. Purification by
medium pressure (30% EtOAc in hexanes to EtOAc) provided
5-(3-{2S-[4-(4-fluoro-3-methyl-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl-
}-propyl)-thiophene-2-carboxylic acid methyl ester (114 mg).
.sup.1H NMR (CDCl.sub.3) .delta. 7.60 (d, 1H), 6.97 (d, 1H), 6.93
(d, 2H), 6.79 (d, 1H), 3.82 (s, 3H), 3.63 (m, 1H), 3.60 (s, 2H),
3.50 (m, 1H), 2.93 (m, 1H), 2.79 (t, 2H), 2.42 (m, 2H), 2.33-2.21
(m, 5H), 2.02-1.78 (m, 4H), 1.50 (m, 2H); MS 446.1 (M+1).
Step C:
5-(3-{2S-[4-(4-Fluoro-3-methyl-phenyl)-3-hydroxy-butyl]-5-oxo-pyr-
rolidin-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester.
Analogous to the procedure described for Compound 2B, Step C,
5-(3-{2S-[4-(4-fluoro-3-methyl-phenyl)-3-oxo-butyl]-5-oxo-pyrrolidin-1-yl-
}-propyl)-thiophene-2-carboxylic acid methyl ester (114 mg, 0.256
mmol) was reduced with NaBH.sub.4 (5 mg, 0.132 mmol) in MeOH (10
mL) at 0.degree. C. to room temperature over 2.5 h. Purification by
medium pressure chromatography (1:1 hexanes:EtOAc to EtOAc)
provided
5-(3-{2S-[4-(4-fluoro-3-methyl-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (80 mg).
.sup.1H NMR (CDCl.sub.3) .delta. 7.59 (d, 1H), 6.98 (d, 1H), 6.93
(m, 2H), 6.80 (d, 1H), 3.81 (s, 3H), 3.74 (m, 1H), 3.60 (m, 2H),
2.99 (m, 1H), 2.82 (t, 2H), 2.72 (m, 1H), 2.54 (m, 1H), 2.33 (m,
2H), 2.22 (s, 3H), 2.08 (m, 1H), 1.96-1.32 (m, 8H); MS 448.1
(M+1).
Step D:
5-(3-{2S-[4-(4-Fluoro-3-methyl-phenyl)-3-hydroxy-butyl]-5-oxo-pyr-
rolidin-1-yl}-propyl)-thiophene-2-carboxylic acid. Analogous to the
procedure described for Compound 2A, Step E,
5-(3-{2S-[4-(4-fluoro-3-methyl-phenyl)-3-hydroxy-butyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (80 mg,
0.179 mmol) was hydrolyzed with 2N NaOH in MeOH (6 mL) over 18 h to
provide the title compound of Example 3J (56 mg). .sup.1H NMR
(CD.sub.3OD) .delta. 7.58 (d, 1H), 7.08-6.98 (m, 2H), 6.90 (m, 2H),
3.69 (m, 2H), 3.55 (m, 1H), 3.04 (m, 1H), 2.84 (t, 2H), 2.67 (m,
2H), 2.31 (m, 2H), 2.21 (s, 3H), 2.11 (m, 1H), 1.98-1.27 (m, 7H);
MS 432.4 (M-1).
Compound 3K
5-{3-[2S-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-thiophe-
ne-2-carboxylic acid
Step A:
5-{3-[2-oxo-5R-(3-oxo-4-phenyl-but-1-enyl)-pyrrolidin-1-yl]-propy-
l}-thiophene-2-carboxylic acid methyl ester. Analogous to the
procedure described for Compound 2A, Step B, the anion derived from
(2-oxo-3-phenyl-propyl)-phosphonic acid dimethyl ester (543 mg,
2.24 mmol) and NaH (60% by weight in oil, 94 mg, 2.35 mmol) was
reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxylic
acid methyl ester (assumed 2.36 mmol) over 18 h. Purification by
medium pressure chromatography (20% EtOAc in hexanes to 70% EtOAc
in hexanes) provided
5-{3-[2-oxo-5R-(3-oxo-4-phenyl-but-1-enyl)-pyrrolidin-1-yl]-prop-
yl}-thiophene-2-carboxylic acid methyl ester (315 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.61 (d, 1H), 7.34-7.15 (m, 5H), 6.77 (m, 1H),
6.61 (dd, 1H), 6.19 (d, 1H), 4.12 (m, 1H), 3.85 (s, 3H), 3.82 (s,
2H), 3.54 (m, 1H), 2.81 (m, 3H), 2.37 (m, 2H), 2.20 (m, 1H), 1.78
(m, 3H); MS 411.8 (M+1); 409.7 (M-1).
Step B:
5-{3-[2-Oxo-5S-(3-oxo-4-phenyl-butyl)-pyrrolidin-1-yl]-propyl}-th-
iophene-2-carboxylic acid methyl ester. Analogous to the procedure
described for Compound 2A, Step D,
5-{3-[2-oxo-5R-(3-oxo-4-phenyl-but-1-enyl)-pyrrolidin-1-yl]-propyl}-thiop-
hene-2-carboxylic acid methyl ester (305 mg, 0.741 mmol) was
hydrogenated in MeOH (30 mL) in the presence of 10% palladium on
carbon (100 mg) at 50 psi for 1.5 h. Purification by medium
pressure (1:1 hexanes:EtOAc to EtOAc) provided
5-{3-[2-oxo-5S-(3-oxo-4-phenyl-butyl)-pyrrolidin-1-yl]-propyl}-thiophene--
2-carboxylic acid methyl ester (235 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 7.62 (d, 1H), 7.35-7.18 (m, 5H), 6.81 (d, 1H), 3.84 (s,
3H), 3.69 (s, 2H), 3.62 (m, 1H), 3.48 (m, 1H), 2.94 (m, 1H), 2.80
(t, 2H), 2.43 (m, 2H), 2.26 (m, 2H), 2.04-1.78 (m, 4H), 1.48 (m,
2H); MS 414.1 (M+1).
Step C:
5-{3-[2S-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl-
}-thiophene-2-carboxylic acid methyl ester. Analogous to the
procedure described for Compound 2B, Step C,
5-{3-[2-oxo-5S-(3-oxo-4-phenyl-butyl)-pyrrolidin-1-yl]-propyl}-thiophene--
2-carboxylic acid methyl ester (235 mg, 0.569 mmol) was reduced
with NaBH.sub.4 (11 mg, 0.284 mmol) in MeOH (7 mL) at 0.degree. C.
to room temperature over 2 h. Purification by medium pressure
chromatography (30% EtOAc in hexanes to EtOAc) provided
5-{3-[2S-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-thioph-
ene-2-carboxylic acid methyl ester (177 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.70 (d, 1H), 7.32-7.16 (m, 5H), 6.79 (d, 1H),
3.80 (m, 4H), 3.60 (m, 2H), 2.99 (m, 1H), 2.80 (m, 3H), 2.62 (m,
1H), 2.32 (m, 2H), 2.09 (m, 1H), 1.97-1.32 (m, 8H); MS 416.0
(M+1).
Step D:
5-{3-[2S-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl-
}-thiophene-2-carboxylic acid. Analogous to the procedure described
for Compound 2A, Step E,
5-{3-[2S-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-thioph-
ene-2-carboxylic acid methyl ester (177 mg, 0.426 mmol) was
hydrolyzed with 2N NaOH in MeOH (7 mL) over 18 h to provide the
title compound of Example 3K (132 mg). .sup.1H NMR (CD.sub.3OD)
.delta. 7.57 (m, 1H), 7.26-7.14 (m, 5H), 6.88 (d, 1H), 3.75 (m,
1H), 3.64 (m, 1H), 3.54 (m, 1H), 3.00 (m, 1H), 2.82 (t, 2H), 2.71
(m, 2H), 2.28 (m, 2H), 2.08 (m, 1H), 1.96-1.26 (m, 7H); MS 402.2
(M+1), 400.4 (M-1).
Compound 3L
5-(3-{2S-[4-(3-Chloro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pro-
pyl)-thiophene-2-carboxylic acid
Step A:
5-(3-{2R-[4-(3-Chloro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous
to the procedure described for Compound 2C, Step D, the anion
derived from [3-(3-chloro-phenyl)-2-oxo-propyl]-phosphonic acid
dimethyl ester (3.68 g, 13.3 mmol) and NaH (60% by weight in oil,
533 mg, 14.5 mmol) was reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxylic
acid methyl ester (assumed 12.1 mmol) over 24 h. Purification by
medium pressure chromatography (15% acetone in toluene to 20%
acetone in toluene) provided
5-(3-{2R-[4-(3-chloro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid methyl ester (2.63 g). .sup.1H
NMR (CDCl.sub.3) .delta. 7.59 (d, 1H), 7.23 (m, 2H), 7.16 (s, 1H),
7.04 (m, 1H), 6.76 (d, 1H), 6.60 (dd, 1H), 6.17 (d, 1H), 4.12 (m,
1H), 3.82 (s, 3H), 3.78 (s, 2H), 3.56 (m, 1H), 2.87-2.75 (m, 3H),
2.45-2.28 (m, 2H), 2.21 (m, 1H), 1.78 (m, 3H).
Step B:
5-(3-{2R-[4-(3-Chloro-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrol-
idin-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. To a
solution of
5-(3-{2R-[4-(3-chloro-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl}-
-propyl)-thiophene-2-carboxylic acid methyl ester (2.63 g, 5.91
mmol) and (R)-2-methyl-CBS-oxazaborolidine (1M in toluene, 5.9 mL,
5.9 mmol in CH.sub.2Cl.sub.2 (140 mL) at -45.degree. C. was added
catecholborane (1M in THF, 17.7 mL, 17.7 mmol) dropwise. The
reaction mixture was stirred for 18 h and MeOH was added. After
stirring for 18 h, the volatiles were removed in vacuo and
CH.sub.2Cl.sub.2 was added. The organic solution was washed with
cold 1N NaOH (3 times), 1N HCl, water and brine. The organic
solution was dried (MgSO.sub.4), filtered and concentrated.
Purification by medium pressure chromatography (1:1 hexanes:EtOAc
to 80% EtOAc in hexanes) provided
5-(3-{2R-[4-(3-chloro-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrolidin-1-y-
l}-propyl)-thiophene-2-carboxylic acid methyl ester (870 mg) as an
approximate 10:1 ratio of 3S:3R alcohol diastereomers by .sup.1H
NMR. .sup.1H NMR (CDCl.sub.3) .delta. 7.61 (d, 1H), 7.21 (m, 3H),
7.07 (m, 1H), 6.80 (d, 1H), 5.68 (dd, 1H), 5.45 (dd, 1H), 4.36 (m,
1H), 4.01 (m, 1H), 3.82 (s, 3H), 3.51 (m, 1H), 2.84-2.76 (m, 5H),
2.44-2.28 (m, 2H), 2.18 (m, 1H), 1.86-1.56 (m, 4H).
Step C:
5-(3-{2S-[4-(3-Chloro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester. Analogous
to the procedure described for Compound 2A, Step D, a mixture of
5-(3-{2R-[4-(3-chloro-phenyl)-3S-hydroxy-but-1-enyl]-5-oxo-pyrrolidin-1-y-
l}-propyl)-thiophene-2-carboxylic acid methyl ester (850 mg) and
10% palladium on carbon (100 mg) in MeOH (50 mL) was hydrogenated
on a Parr shaker at 50 psi for 3 h. The hydrogenation was repeated
using 100 mg of 10% palladium on carbon for 6 h. Purification by
medium pressure chromatography (1:1 hexanes:EtOAc to EtOAc)
provided
5-(3-{2S-[4-(3-chloro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid methyl ester (504 mg). .sup.1H
NMR (CDCl.sub.3) .delta. 7.61 (d, 1H), 7.23 (m, 3H), 7.08 (m, 1H),
6.82 (d, 1H), 3.83 (s, 3H), 3.81 (m, 1H), 3.62 (m, 2H), 3.01 (m,
1H), 2.84 (t, 2H), 2.77 (m, 1H), 2.65 (m, 1H), 2.35 (m, 2H), 2.10
(m, 1H), 1.97-1.43 (m, 8H).
Step D:
5-(3-{2S-[4-(3-Chloro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid. Analogous to the
procedure described for Compound 2A, Step E,
5-(3-{2S-[4-(3-chloro-phenyl)-3R-hydroxy-butyl]-5-oxo-pyrrolidin-1-yl}-pr-
opyl)-thiophene-2-carboxylic acid methyl ester (504 mg) was
hydrolyzed with 2N NaOH in MeOH (20 mL) at 50.degree. C. over 4 h
to provide the title compound of Example 3L (338.6 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 7.68 (d, 1H), 7.22 (m, 3H), 7.08 (m, 1H), 6.84
(d, 1H), 3.80 (m, 1H), 3.64 (m, 2H), 3.01 (m, 1H), 2.82 (m, 4H),
2.64 (m, 1H), 2.38 (m, 2H), 2.12 (m, 1H), 1.92 (m, 3H), 1.66 (m,
1H), 1.57-1.19 (m, 3H). MS 436.1 (M+1), 434.2 (M-1).
Compound 3M
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin--
1-yl}-propyl)-thiophene-2-carboxylic acid
Step A:
5-(3-{2-Oxo-5R-[3-oxo-4-(3-trifluoromethyl-phenyl)-but-1-enyl]-py-
rrolidin-}-propyl)-thiophene-2-carboxylic acid methyl ester.
Analogous to the procedure described for Compound 2A, Step B, the
anion derived from
[2-oxo-3-(3-trifluoromethyl-phenyl)-propyl]-phosphonic acid
dimethyl ester (5.026 g, 17.0 mmol) and NaH (60% by weight in oil,
750 mg, 18.8 mmol) was reacted with
5-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxylic
acid methyl ester (assumed 18.8 mmol) over 24 h. Purification by
medium pressure chromatography (15% acetone in toluene to 20%
acetone in toluene) provided
5-(3-{2-oxo-5R-[3-oxo-4-(3-trifluoromethyl-phenyl)-but-1-enyl]-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (4.02 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.61 (d, 1H), 7.54 (d, 1H), 7.45
(m, 2H), 7.37 (d, 1H), 6.79 (d, 1H), 6.66 (dd, 1H), 6.20 (d, 1H),
4.16 (m, 1H), 3.90 (s, 2H), 3.84 (s, 3H), 3.60 (m, 1H), 2.89-2.78
(m, 3H), 2.48-2.31 (m, 2H), 2.23 (m, 1H), 1.82 (m, 3H).
Step B:
5-(3-{2R-[3S-Hydroxy-4-(3-trifluoromethyl-phenyl)-but-1-enyl]-5-o-
xo-pyrrolidin-1-yl}-propyl)-thiophene-2-carboxylic acid methyl
ester. Analogous to the procedure described for Compound 2A, Step
C,
5-(3-{2-oxo-5R-[3-oxo-4-(3-trifluoromethyl-phenyl)-but-1-enyl]-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (2.63 g,
5.91 mmol) was reduced with catecholborane (1M in THF, 18.8 mL,
18.8 mmol) in the presence of (R)-2-methyl-CBS-oxazaborolidine (1M
in toluene, 0.94 mL, 0.94 mmol) at -45.degree. C. over 18 h. The
reaction was quenched by addition of 1N HCl and the mixture was
stirred for 40 minutes. The organic solution was washed
consecutively with ice cold 1N NaOH (3 times), 1N HCl (1 time),
water (1 time), and brine. The organic solution was dried
(MgSO.sub.4), filtered, and concentrated. Purification by medium
pressure chromatography (10% acetone in toluene to 20% acetone in
toluene) provided
5-(3-{2R-[3S-hydroxy-4-(3-trifluoromethyl-phenyl)-but-1-enyl]-5-oxo-pyrro-
lidin-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (3 g)
as an approximate 4:1 ratio of 3S:3R alcohol diastereomers by
.sup.1H NMR. .sup.1H NMR (CDCl.sub.3) .delta. 7.60 (d, 1H), 7.50
(d, 1H), 7.41 (m, 3H), 6.79 (d, 1H), 5.70 (dd, 1H), 5.48 (dd, 1H),
4.41 (m, 1H), 4.00 (m, 1H), 3.81 (s, 3H), 3.50 (m, 1H), 2.86-2.77
(m, 5H), 2.42-2.26 (m, 2H), 2.16 (m, 1H), 1.81 (m, 2H), 1.72-1.54
(m, 2H).
Step C:
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-py-
rrolidin-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester.
Analogous to the procedure described for Compound 2A, Step D, a
mixture of
5-(3-{2R-[3S-hydroxy-4-(3-trifluoromethyl-phenyl)-but-1-enyl]-5-oxo-pyrro-
lidin-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (3 g)
and 10% palladium on carbon (400 mg) in MeOH (70 mL) was
hydrogenated on a Parr shaker at 50 psi for 16 h. Purification by
medium pressure chromatography (20% EtOAc in hexanes to 70% EtOAc
in hexanes) provided
5-(3-{2S-[3R-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (2.26 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.61 (d, 1H), 7.52-7.38 (m, 4H),
6.81 (d, 1H), 3.83 (m, 4H), 3.63 (m, 2H), 3.00 (m, 1H), 2.85 (m,
3H), 2.74 (m, 1H), 2.34 (m, 2H), 2.10 (m, 1H), 1.98-1.45 (m,
08H).
Step D:
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-py-
rrolidin-1-yl}-propyl)-thiophene-2-carboxylic acid. Analogous to
the procedure described for Compound 2A, Step E,
5-(3-{2S-[3R-hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid methyl ester (625 mg)
was hydrolyzed with 2N NaOH in MeOH (20 mL) at room temperature
over 24 h to provide the title compound of Example 3M (599 mg).
.sup.1H NMR (CDCl.sub.3) .delta. 7.67 (d, 1H), 7.51-7.38 (m, 4H),
6.84 (d, 1H), 3.85 (m, 1H), 3.63 (m, 2H), 3.02 (m, 1H), 2.85 (m,
3H), 2.75 (m, 1H), 2.37 (m, 2H), 2.11 (m, 1H), 2.00-1.45 (m, 8H);
MS 470.2 (M+1), 468.2 (M-1).
The sodium salt of Compound 3M was prepared by addition of sodium
bicarbonate (1.0 equivalent) to a solution of Compound 3M (1.0
equivalent) in an ethanol/water mixture. The mixture was stirred
and then was concentrated in vacuo to dryness to provide Compound
3M as the sodium salt.
Compound 4A
5S-(3-Hydroxy-4-naphthalen-2-yl-butyl)-1-[6-(2H-tetrazol-5-yl)-hexyl]-pyrr-
olidin-2-one
Step A: 7-(2R-Formyl-5-oxo-pyrrolidin-1-yl)-heptanenitrile.
Analogous to the procedure described for Compound 2A, Step A,
7-(2R-hydroxymethyl-5-oxo-pyrrolidin-1-yl)-heptanenitrile (150 mg,
0.67 mmol) was oxidized to generate
7-(2R-formyl-5-oxo-pyrrolidin-1-yl)-heptanenitrile which was used
in Step B without further purification.
Step B:
7-[2R-(4-Naphthalen-2-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-
-heptanenitrile. Analogous to the procedure described for Compound
2A, Step B, the anion derived from
(3-naphthalen-2-yl-2-oxo-propyl)-phosphonic acid dimethyl ester
(196 mg, 0.67 mmol) and NaH (60% by weight in oil, 27 mg, 0.67
mmol) was reacted with
7-(2R-formyl-5-oxo-pyrrolidin-1-yl)-heptanenitrile (assumed 0.67
mmol) over 19 h. Purification by medium pressure chromatography
(1:1 hexanes:EtOAc to EtOAc) provided
7-[2R-(4-naphthalen-2-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-heptane-
nitrile (74 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.79 (m, 3H),
7.67 (m, 1H), 7.46 (m, 2H), 7.30 (d, 1H), 6.65 (dd, 1H), 6.25 (d,
1H), 4.10 (m, 1H), 3.99 (s, 2H), 3.42 (m, 1H), 2.66 (m, 1H), 2.37
(m, 2H), 2.22 (m, 3H), 1.76 (m, 1H), 1.52 (m, 2H), 1.29 (m, 4H),
1.10 (m, 2H); MS 389.1 (M+1), 387.0 (M-1).
Step C:
7-[2S-(4-Naphthalen-2-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-hept-
anenitrile. Analogous to the procedure described for Compound 2A,
Step D,
7-[2R-(4-naphthalen-2-yl-3-oxo-but-1-enyl)-5-oxo-pyrrolidin-1-yl]-heptane-
nitrile (74 mg, 0.19 mmol) was hydrogenated in EtOH (30 mL) in the
presence of 10% palladium on carbon (50 mg) at 50 psi for 3 h.
Purification by medium pressure (1:1 hexanes:EtOAc to EtOAc)
provided
7-[2S-(4-naphthalen-2-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-heptanenitri-
le (45 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.80 (m, 3H), 7.66 (s,
1H), 7.47 (m, 2H), 7.30 (d, 1H), 3.85 (s, 2H), 3.51 (m, 2H), 2.81
(m, 1H), 2.48 (m, 2H), 2.28 (m, 4H), 1.98 (m, 2H), 1.62 (m, 4H),
1.44 (m, 4H), 1.22 (m, 2H); MS 391.4 (M+1), 389.3 (M-1).
Step D:
7-[2S-(3-Hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]--
heptanenitrile. Analogous to the procedure described for Compound
2B, Step C,
7-[2S-(4-naphthalen-2-yl-3-oxo-butyl)-5-oxo-pyrrolidin-1-yl]-heptaneni-
trile (42 mg, 0.108 mmol) was reduced with NaBH.sub.4 (4 mg, 0.11
mmol) in EtOH (20 mL) at room temperature for 3 h to provide
7-[2S-(3-hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-heptanen-
itrile (40 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.80 (m, 3H), 7.65
(m, 1H), 7.46 (m, 2H), 7.33 (d, 1H), 3.92 (m, 1H), 3.59 (m, 2H),
3.03-2.78 (m, 3H), 2.35 (m, 4H), 2.12 (m, 1H), 1.81 (m, 1H),
1.68-1.40 (m, 11H), 1.28 (m, 2H); MS 393.1 (M+1).
Step E:
5S-(3-Hydroxy-4-naphthalen-2-yl-butyl)-1-[6-(2H-tetrazol-5-yl)-he-
xyl]-pyrrolidin-2-one. A solution of
7-[2S-(3-hydroxy-4-naphthalen-2-yl-butyl)-5-oxo-pyrrolidin-1-yl]-heptanen-
itrile (39 mg, 0.0994 mmol), azidotrimethylsilane (150 mg, 1.30
mmol), and dibutyltin oxide (25 mg, 0.10 mmol) in toluene (15 mL)
was heated under reflux for 19 h. The reaction mixture was cooled
and was acidified to pH of 2 with 1N HCl (5 mL). The volatiles were
removed in vacuo and the aqueous solution was washed with EtOAc
(4.times.10 mL). The organic solutions were combined, dried
(MgSO.sub.4), filtered and concentrated. The residue was purified
by preparative thin layer chromatography (9:1 EtOAc:MeOH) to
provide
5S-(3-hydroxy-4-naphthalen-2-yl-butyl)-1-[6-(2H-tetrazol-5-yl)-hexyl]-pyr-
rolidin-2-one (11 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.79 (m,
3H), 7.65 (m, 1H), 7.45 (m, 2H), 7.32 (m, 1H), 3.94 (m, 1H), 3.66
(m, 1H), 3.52 (m, 1H), 3.03-2.83 (m, 5H), 2.44 (m, 2H), 2.18 (m,
1H), 1.87-1.20 (m, 14H); MS 436.1 (M+1), 435.2 (M-1).
Compound 4B
5S-[3R-Hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-1-[6-(2H-tetrazol-5-yl)-h-
exyl]-pyrrolidin-2-one
Step A:
7-{2R-[4-(3-Methoxymethyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrroli-
din-1-yl}-heptanenitrile. Analogous to the procedure described for
Compound 2A, Step B, the anion derived from
[3-(3-methoxymethyl-phenyl)-2-oxo-propyl]-phosphonic acid diethyl
ester (2.87 g, 9.13 mmol) and NaH (60% in oil, 446 mg, 11.2 mmol)
was reacted with 7-(2R-formyl-5-oxo-pyrrolidin-1-yl)-heptanenitrile
(assumed 11.15 mmol) over 24 h. Purification by medium pressure
chromatography (1:1 hexanes:EtOAc to EtOAc to 1% MeOH in
CH.sub.2Cl.sub.2 to 3% MeOH in CH.sub.2Cl.sub.2) provided
7-{2R-[4-(3-methoxymethyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl-
}-heptanenitrile (2.06 g). .sup.1H NMR (CDCl.sub.3) .delta.0 7.29
(m, 1H), 7.22 (m, 1H), 7.16 (s, 1H), 7.10 (m, 1H), 6.62 (dd, 1H),
6.20 (d, 1H), 4.41 (s, 2H), 4.12 (m, 1H), 3.82 (s, 2H), 3.49 (m,
1H), 3.37 (s, 3H), 2.72 (m, 1H), 2.43-2.20 (m, 5H), 1.76 (m, 1H),
1.60 (m, 2H), 1.40 (m, 4H), 1.24 (m, 2H)
Step B:
7-{2R-[3S-Hydroxy-4-(3-methoxymethyl-phenyl)-but-1-enyl]-5-oxo-py-
rrolidin-1-yl}-heptanenitrile. To a solution of
7-{2R-[4-(3-methoxymethyl-phenyl)-3-oxo-but-1-enyl]-5-oxo-pyrrolidin-1-yl-
}-heptanenitrile (2.06 g, 5.39 mmol) and
(R)-2-methyl-CBS-oxazaborolidine (1M in toluene, 0.81 mL, 0.81
mmol) in CH.sub.2Cl.sub.2 (200 mL) at -45.degree. C. was added
catecholborane (1M in THF, 16.2 mL, 16.2 mmol) dropwise. The
reaction mixture was stirred at -45.degree. C. for 24 h and 1N HCl
was added. The reaction mixture was stirred at room temperature for
1 h and the layers were separated. The aqueous solution was washed
with CH.sub.2Cl.sub.2 (2 times) and the organic solutions were
combined, washed with cold 1N NaOH followed by brine 2 times. The
organic solution was dried (MgSO.sub.4), filtered and concentrated.
Purification by medium pressure chromatography (1:1 hexanes:EtOAc
to EtOAc to 1% MeOH in CH.sub.2Cl.sub.2 to 3% MeOH in
CH.sub.2Cl.sub.2) provided
7-{2R-[3S-hydroxy-4-(3-methoxymethyl-phenyl)-but-1-enyl]-5-oxo-pyrrolidin-
-1-yl}-heptanenitrile (2.07 g) as an approximate 2:1 mixture of
3S:3R alcohol diastereomers by .sup.1H NMR. .sup.1H NMR
(CDCl.sub.3) .delta. 7.30-7.09 (m, 4H), 5.71 (m, 1H), 5.46 (m, 1H),
4.41 (s, 2H), 4.38 (m, 1H), 4.00 (m, 1H), 3.45 (m, 1H), 3.38 (s,
3H), 2.88-2.68 (m, 3H), 2.31 (m, 4H), 2.17 (m, 1H), 1.70-1.21 (m,
10H).
Step C:
7-{2S-[3R-Hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-5-oxo-pyrroli-
din-1-yl}-heptanenitrile. Analogous to the procedure described for
Compound 2A, Step D,
7-{2R-[3S-hydroxy-4-(3-methoxymethyl-phenyl)-but-1-enyl]-5-oxo-pyrrolidin-
-1-yl}-heptanenitrile (2.07 g, 5.39 mmol) in EtOH (100 mL) was
hydrogenated in the presence of 10% palladium on carbon (200 mg) at
50 psi for 24 h on a Parr shaker. Purification by medium pressure
chromatography (1:1 hexanes:EtOAc to 2:1 EtOAc:hexanes to EtOAc to
2% MeOH in CH.sub.2Cl.sub.2 to 5% MeOH in CH.sub.2Cl.sub.2 to 10%
MeOH in CH.sub.2Cl.sub.2) provided
7-{2S-[3R-hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl-
}-heptanenitrile (1.28 g). .sup.1H NMR (CDCl.sub.3) .delta.
7.30-7.10 (m, 4H), 4.41 (s, 2H), 3.82 (m, 1H), 3.57 (m, 2H), 3.38
(s, 3H), 2.89 (m, 2H), 2.66 (m, 1H), 2.32 (m, 4H), 2.10 (m, 1H),
1.77 (m, 1H), 1.66-1.40 (m, 11H), 1.29 (m, 2H).
Step D:
5S-[3R-Hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-1-[6-(2H-tetrazo-
l-5-yl)-hexyl]-pyrrolidin-2-one. Analogous to the procedure
described for Compound 4A, Step E,
7-{2S-[3R-hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-5-oxo-pyrrolidin-1-yl-
}-heptanenitrile (1.28 g, 3.31 mmol) was reacted with
azidotrimethylsilane (0.90 mL, 6.78 mmol) and dibutyltin oxide (128
mg, 0.514 mmol) in toluene (68 mL) heated under reflux for 24 h.
Additional azidotrimethylsilane (1.8 mL, 13.56 mmol) and dibutyltin
oxide (256 mg, 1.03 mmol) were added and the reaction mixture was
continued under reflux for 3 days. Purification by medium pressure
chromatography (CH.sub.2Cl.sub.2 to 2% MeOH in CH.sub.2Cl.sub.2 to
4% MeOH in CH.sub.2Cl.sub.2 to 6% MeOH in CH.sub.2Cl.sub.2 to 10%
MeOH in CH.sub.2Cl.sub.2) provided
5S-[3R-hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-1-[6-(2H-tetrazol-5-yl)--
hexyl]-pyrrolidin-2-one (619.5 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 7.30-7.11 (m, 4H), 4.42 (s, 2H), 3.87 (m, 1H), 3.64 (m,
1H), 3.52 (m, 1H), 3.39 (s, 3H), 2.99-2.67 (m, 5H), 2.42 (m, 2H),
2.16 (m, 1H), 1.87-1.25 (m, 14H).
Step E: Sodium salt of
5S-[3R-Hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-1-[6-(2H-tetrazol-5-yl)--
hexyl]-pyrrolidin-2-one. Analogous to the procedure described for
Compound 2C, Step D, treatment of
5S-[3R-hydroxy-4-(3-methoxymethyl-phenyl)-butyl]-1-[6-(2H-tetrazol-5-yl)--
hexyl]-pyrrolidin-2-one (619.5 mg, 1.44 mmol) with NaHCO.sub.3 (121
mg, 1.44 mmol) provided the sodium salt of the title compound,
Compound 4B (628.3 mg). .sup.1H NMR (CD.sub.3OD) .delta. 7.20 (m,
4H), 3.79 (m, 1H), 3.64 (m, 1H), 3.50 (m, 1H), 2.97-2.69 (m, 5H),
2.29 (m, 2H), 2.10 (m, 1H), 1.81-1.28 (m, 14H).
Compound 5A
2-{3-[2S-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-thiazol-
e-4-carboxylic acid
Step A:
2-{3-[2-Oxo-5R-(3-oxo-4-phenyl-but-1-enyl)-pyrrolidin-1-yl]-propy-
l}-thiazole-4-carboxylic acid ethyl ester. Analogous to the
procedure described for Compound 2A, Step B, the anion derived from
(2-oxo-3-phenyl-propyl)-phosphonic acid dimethyl ester (105 mg,
0.434 mmol) and NaH (60% by weight in oil, 17 mg, 0.434 mmol) was
reacted with
2-[3-(2R-formyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiazole-4-carboxylic
acid ethyl ester (prepared from
2-[3-(2R-hydroxymethyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiazole-4-carboxyl-
ic acid ethyl ester analogous to the procedure described for
Compound 2A, Step A, assumed 0.359 mmol) over 17 h. Purification by
medium pressure chromatography (1:1 hexanes:EtOAc to EtOAc)
provided
2-{3-[2-oxo-5R-(3-oxo-4-phenyl-but-1-enyl)-pyrrolidin-1-yl]-propyl}-thiaz-
ole-4-carboxylic acid ethyl ester (59 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 8.03 (s, 1H), 7.33-7.17 (m, 5H), 6.61 (dd, 1H), 6.20 (d,
1H), 4.40 (q, 2H), 4.19 (m, 1H), 3.82 (s, 2H), 3.60 (m, 1H), 2.98
(m, 2H), 2.80 (m, 1H), 2.44-2.15 (m, 3H), 1.94 (m, 2H), 1.75 (m,
1H), 1.38 (t, 3H); MS 427.0 (M+1), 424.9 (M-1).
Step B:
2-{3-[2-Oxo-5S-(3-oxo-4-phenyl-butyl)-pyrrolidin-1-yl]-propyl}-th-
iazole-4-carboxylic acid ethyl ester. Analogous to the procedure
described for Compound 2A, Step D,
2-{3-[2-oxo-5R-(3-oxo-4-phenyl-but-1-enyl)-pyrrolidin-1-yl]-propyl}-thiaz-
ole-4-carboxylic acid ethyl ester (23 mg, 0.0539 mmol) was
hydrogenated in EtOH (15 mL) in the presence of 10% palladium on
carbon (15 mg) at 50 psi for 3 h. Purification by preparative thin
layer chromatography (1:1 hexanes:EtOAc) (2 times) provided
2-{3-[2-oxo-5S-(3-oxo-4-phenyl-butyl)-pyrrolidin-1-yl]-propyl}-thiazole-4-
-carboxylic acid ethyl ester (19 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 8.03 (s, 1H), 7.34-7.17 (m, 5H), 4.39 (q, 2H), 3.68 (s,
2H), 3.65 (m, 1H), 3.53 (m, 1H), 2.98 (m, 3H), 2.43 (t, 2H), 2.26
(m, 2H), 1.98 (m, 4H), 1.49 (m, 2H), 1.37 (t, 3H) MS 429.0
(M+1).
Step C:
2-{3-[2S-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl-
}-thiazole-4-carboxylic acid ethyl ester. Analogous to the
procedure described for Compound 2B, Step C,
2-{3-[2-oxo-5S-(3-oxo-4-phenyl-butyl)-pyrrolidin-1-yl]-propyl}-thiazole-4-
-carboxylic acid ethyl ester (34 mg, 0.0793 mmol) was reduced with
NaBH.sub.4 (3 mg, 0.079 mmol) in EtOH (10 mL) at room temperature
for 2 h. Purification by preparative thin layer chromatography
(EtOAc) provided
2-{3-[2S-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-thiazo-
le-4-carboxylic acid ethyl ester (18 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 8.02 (m, 1H), 7.33-7.18 (m, 5H), 4.38 (q, 2H), 3.82 (m,
1H), 3.65 (m, 2H), 3.06 (m, 3H), 2.80 (m, 1H), 2.67 (m, 1H), 2.32
(m, 2H), 2.09 (m, 2H), 1.98 (m, 2H), 1.82 (m, 1H), 1.68-1.42 (m,
4H), 1.37 (t, 3MS 431.1 (M+1).
Step D:
2-{3-[2S-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl-
}-thiazole-4-carboxylic acid. Analogous to the procedure described
for Compound 2A, Step E,
2-{3-[2S-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-thiazo-
le-4-carboxylic acid ethyl ester (18 mg, 0.042 mmol) was hydrolyzed
with 1 N NaOH (0.06 mL) in MeOH (5 mL) heated under reflux for 3 h
to provide the title compound of Example 5A (8 mg). .sup.1H NMR
(CDCl.sub.3) .delta. 8.01 (s, 1H), 7.33-7.18 (m, 5H), 3.83 (m, 1H),
3.66 (m, 2H), 3.09 (m, 1H), 3.02 (t, 2H), 2.81 (m, 1H), 2.68 (m,
1H), 2.35 (m, 2H), 2.06 (m, 4H), 1.82 (m, 1H), 1.69-1.38 (m, 4H);
MS 403.0 (M+1), 401.0 (M-1).
Step E: Sodium salt of
2-{3-[2S-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-thiazo-
le-4-carboxylic acid. The sodium salt of the title compound,
Compound 5A was prepared analogous to the procedure described for
Compound 2B, Step E. .sup.1H NMR (CDCl.sub.3) .delta. 7.58 (s, 1H),
7.25-7.14 (m, 5H), 3.75 (m, 1H), 3.36 (m, 2H), 2.78 (m, 1H), 2.61
(m, 3H), 2.16-1.20 (m, 12H).
Compound 5B
5-(3-Hydroxy-4-phenyl-butyl)-1-{3-[4-(2H-tetrazol-5-yl)-phenyl]-propyl}-py-
rrolidin-2-one
Step A:
4-(3-{2-[3-(tert-Butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-5-oxo-
-pyrrolidin-1-yl}-propyl)-benzonitrile. Analogous to the procedure
described for Compound 1A, Step D, the anion derived from
5-[3-(tert-butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-pyrrolidin-2-one
(262.8 mg, 0.756 mmol) and NaHMDS (0.83 mL, 0.83 mmol) was reacted
with 4-(3-bromo-propyl)-benzonitrile (186 mg, 0.832 mmol) at
70.degree. C. for 24 h. Purification by medium pressure
chromatography (5:1 hexanes:EtOAc to 1:1 hexanes:EtOAc to 1% MeOH
in CH.sub.2Cl.sub.2 to 5% MeOH in CH.sub.2Cl.sub.2) provided
4-(3-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-5-oxo-pyrroli-
din-1-yl}-propyl)-benzonitrile (257.6 mg). .sup.1H NMR (CDCl.sub.3)
.delta. 7.56 (m, 2H), 7.26 (m, 5H), 7.13 (m, 2H), 3.85 (m, 1H),
3.62 (m, 1H), 3.48 (m, 1H), 2.93 (m, 1H), 2.82-2.60 (m, 4H), 2.29
(m, 2H), 1.88-1.25 (m, 7H); MS 491.5 (M+1).
Step B:
4-{3-[2-(3-Hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-
-benzonitrile. Analogous to the procedure described for Compound
1A, Step E,
4-(3-{2-[3-(tert-butyl-dimethyl-silanyloxy)-4-phenyl-butyl]-5-oxo-pyrr-
olidin-1-yl}-propyl)-benzonitrile (257.6 mg, 0.525 mmol) was
deprotected with TBAF (1M in THF, 0.79 mL, 0.79 mmol) over 24 h.
Purification by medium pressure chromatography (1:1 EtOAc: hexanes
to EtOAc to 1% MeOH in CH.sub.2Cl.sub.2 to 3% MeOH in
CH.sub.2Cl.sub.2) provided
4-{3-[2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-benzoni-
trile (157.8 mg). .sup.1H NMR (CDCl.sub.3) .delta. 7.56 (m, 2H),
7.26 (m, 7H), 3.80 (m, 1H), 3.67-3.55 (m, 2H), 2.98 (m, 1H), 2.80
(m, 1H), 2.65 (t, 2H), 2.43-2.24 (m, 2H), 2.08 (m, 1H), 1.89-1.33
(m, 9H); MS 375.3 (M-1).
Step C:
5-(3-Hydroxy-4-phenyl-butyl)-1-{3-[4-(2H-tetrazol-5-yl)-phenyl]-p-
ropyl}-pyrrolidin-2-one. Analogous to the procedure described for
Compound 4A, Step E,
4-{3-[2-(3-hydroxy-4-phenyl-butyl)-5-oxo-pyrrolidin-1-yl]-propyl}-benzoni-
trile (157.8 mg, 0.419 mmol) was reacted with azidotrimethylsilane
(0.11 mL, 0.84 mmol) and dibutyltin oxide (20 mg, 0.08 mmol) in
toluene (8.6 mL) heated under reflux for 60 h. Purification by
medium pressure chromatography (CH.sub.2Cl.sub.2 to 2% MeOH in
CH.sub.2Cl.sub.2 to 4% MeOH in CH.sub.2Cl.sub.2 to 6% MeOH in
CH.sub.2Cl.sub.2) provided
5-(3-hydroxy-4-phenyl-butyl)-1-{3-[4-(2H-tetrazol-5-yl)-phenyl]-propyl}-p-
yrrolidin-2-one (144.7 mg). .sup.1H NMR (CDCl.sub.3) .delta. 8.02
(m, 2H), 7.27 (m, 7H), 3.84 (m, 1H), 3.67 (m, 2H), 3.10 (m, 1H),
2.84 (m, 1H), 2.67 (m, 2H), 2.53 (m, 1H), 2.42 (m, 1H), 2.14 (m,
1H), 1.97-1.40 (m, 9H); MS 420.3 (M+1), 418.3 (M-1).
Preparation 1
5-[3-(2R-Hydroxymethyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxyl-
ic acid methyl ester
Step A:
5R-(tert-Butyl-dimethyl-silanyloxymethyl)-1-prop-2-ynyl-pyrrolidi-
n-2-one. To a solution of
5R-(tert-butyl-dimethyl-silanyloxymethyl)-pyrrolidin-2-one
(Tetrahdedron Asymmetry 1996, 7, 2113) (10.24 g, 44.6 mmol) in DMF
(650 mL) at 0.degree. C. was added NaHMDS (1M in THF, 49 mL, 49
mmol) dropwise. The reaction mixture was mechanically stirred at
room temperature for 2 h to yield a thick suspension. The reaction
mixture was cooled to 0.degree. C. and propargyl bromide (80% in
toluene, 5.0 mL, 45 mmol) in DMF (50 mL) was added slowly. The
reaction mixture was stirred at 0.degree. C. for 2 h and at room
temperature for 0.5 h. Aqueous saturated ammonium chloride (700 mL)
and water (300 mL) were added. The solution was washed with EtOAc
(3.times.600 mL). The organic solutions were combined, washed with
water (4.times.300 mL) followed by brine (1.times.300 mL). The
organic solution was dried (Na.sub.2SO.sub.4), filtered and
concentrated. Purification by medium pressure chromatography (10%
EtOAc in hexanes to 25% EtOAc in hexanes) provided
5R-(tert-butyl-dimethyl-silanyloxymethyl)-1-prop-2-ynyl-pyrrolidin-2-one
(9.85 g). .sup.1H NMR (CDCl.sub.3) .delta. 4.58 (dd, 1H), 3.88 (m,
1H), 3.77 (dd, 1H), 3.70 (d, 1H), 3.61 (m, 1H), 2.50-2.28 (m, 2H),
2.18 (m, 1H), 2.10 (m, 1H), 1.86 (m, 1H), 0.87 (s, 9H), 0.05 (s,
6H); MS 268.2 (M+1).
Step B:
5-{3-[2R-(tert-Butyl-dimethyl-silanyloxymethyl)-5-oxo-pyrrolidin--
1-yl]-prop-1-ynyl}-thiophene-2-carboxylic acid methyl ester. A
mixture of
5R-(tert-butyl-dimethyl-silanyloxymethyl)-1-prop-2-ynyl-pyrrolidin-2-one
(8.64 g, 32.3 mmol), 5-bromo-thiophene-2-carboxylic acid methyl
ester (7.5 g, 33.9 mmol), copper (I) iodide, CuI (308 mg, 1.62
mmol), tetrakis(triphenylphosphine)palladium(0) (1.9 g, 1.62 mmol),
triethylamine (5.0 mL, 36 mmol), and CH.sub.3CN (300 mL) was heated
under reflux for 19 h. The reaction mixture was cooled to room
temperature and the volatiles were removed in vacuo. The residue
was dissolved in EtOAc (500 mL) and the organic solution was washed
with water (3.times.200 mL) followed by brine (1.times.200 mL). The
organic solution was dried (Na.sub.2SO.sub.4), filtered and
concentrated. Purification by medium pressure chromatography (10%
EtOAc in hexanes to 25% EtOAc in hexanes) (2 times) provided
5-{3-[2R-(tert-butyl-dimethyl-silanyloxymethyl)-5-oxo-pyrrolidin-1-yl]-pr-
op-1-ynyl}-thiophene-2-carboxylic acid methyl ester (11.42 g).
.sup.1H NMR (CDCl.sub.3) .delta. 7.61 (d, 1H), 7.09 (d, 1H), 4.81
(d, 1H), 3.98 (d, 1H), 3.87 (m, 1H), 3.85 (s, 3H), 3.78 (dd, 1H),
3.63 (dd, 1H), 2.49-2.29 (m, 2H), 2.11 (m, 1H), 1.82 (m, 1H); 0.85
(s, 9H), 0.03 (s, 6H); MS 408.0 (M+1).
Step C:
5-{3-[2R-(tert-Butyl-dimethyl-silanyloxymethyl)-5-oxo-pyrrolidin--
1-yl]-propyl}-thiophene-2-carboxylic acid methyl ester. A mixture
of
5-{3-[2R-(tert-butyl-dimethyl-silanyloxymethyl)-5-oxo-pyrrolidin-1-yl]-pr-
op-1-ynyl}-thiophene-2-carboxylic acid methyl ester (11.4 g, 28
mmol) in EtOH (200 mL) was hydrogenated on a Parr shaker at 50 psi
in the presence of 10% palladium on carbon (1.2 g) for 3 h. The
catalyst was removed by filtration through Celite.RTM.
(diatomaceous earth, Fluka Chemical Corp, Milwaukee, Wis.) with the
aid of EtOH and the organic solution was concentrated in vacuo. The
hydrogenation was repeated using EtOH (200 mL) and 10% palladium on
carbon (1.2 g) at 50 psi for 24 h. Purification by medium pressure
chromatography (25% EtOAc in hexanes to 50% EtOAc in hexanes)
provided
5-{3-[2R-(tert-butyl-dimethyl-silanyloxymethyl)-5-oxo-pyrrolidin-1-yl]-pr-
opyl}-thiophene-2-carboxylic acid methyl ester (10.2 g). .sup.1H
NMR (CDCl.sub.3) .delta. 7.64 (d, 1H), 6.83 (d, 1H), 3.87 (s, 3H),
3.64 (m, 3H), 3.13 (m, 1H), 2.86 (t, 2H), 2.51-2.24 (m, 2H),
2.12-1.78 (m, 4H), 0.88 (s, 9H), 0.04 (s, 6H).
Step D:
5-[3-(2R-Hydroxymethyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-
-carboxylic acid methyl ester. To a solution of
5-{3-[2R-(tert-butyl-dimethyl-silanyloxymethyl)-5-oxo-pyrrolidin-1-yl]-pr-
opyl}-thiophene-2-carboxylic acid methyl ester (1.5 g, 3.64 mmol)
in MeOH (40 mL) was added 1N HCl (18 mL) and the reaction mixture
was stirred for 1.5 h. The volatiles were removed in vacuo and the
aqueous solution was washed with CH.sub.2Cl.sub.2 (3.times.50 mL).
The organic solutions were combined, washed with brine, dried
(MgSO.sub.4), filtered and concentrated. Purification by medium
pressure chromatography (5% MeOH in CH.sub.2Cl.sub.2) provided
5-[3-(2R-hydroxymethyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiophene-2-carboxy-
lic acid methyl ester (689 mg). .sup.1H NMR (CDCl.sub.3) .delta.
7.59 (d, 1H), 6.79 (d, 1H), 3.82 (s, 3H), 3.75 (m, 1 H), 3.62 (m,
3H), 3.07 (m, 1H), 2.82 (t, 2H), 2.44 (m, 1H), 2.26 (m, 2H),
2.09-1.83 (m, 4H); MS 298.2 (M+1).
Preparation 2
7-(2R-Hydroxymethyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid ethyl
ester
Analogous to the procedure described for Preparation 1, Step A, the
anion derived from
5R-(tert-butyl-dimethyl-silanyloxymethyl)-pyrrolidin-2-one (18.83
g, 82.1 mmol) and NaHMDS (1M in THF, 90 mL, 90 mmol) was alkylated
with ethyl 7-bromoheptanoate (16 mL, 82 mmol). The reaction mixture
was stirred at 60.degree. C. for 16 h and was worked-up analogous
to that described for Preparation 1, Step A. The crude residue was
dissolved in MeOH (600 mL) and 1N HCl (300 mL) was added. The
solution was stirred for 3 h and the volatiles were removed in
vacuo. The aqueous solution was diluted with CH.sub.2Cl.sub.2 (300
mL) and the organic solution was washed with water (2.times.75 mL)
followed by brine (1.times.75 mL). The organic solution was dried
(Na.sub.2SO.sub.4), filtered and concentrated. Purification by
medium pressure chromatography (EtOAc) provided
7-(2R-hydroxymethyl-5-oxo-pyrrolidin-1-yl)-heptanoic acid ethyl
ester (21.2 g). .sup.1H NMR (CDCl.sub.3) .delta. 4.12 (q, 2H), 3.80
(dd, 1H), 3.66 (m, 3H), 2.97 (m, 1H), 2.54-2.27 (m, 5H), 2.04 (m,
2H), 1.67-1.28 (m, 8H), 1.26 (t, 3H); MS 272.3 (M+1).
Preparation 3
7-(2R-Hydroxymethyl-5-oxo-pyrrolidin-1-yl)-heptanenitrile
Analogous to the procedure described for Preparation 1, Step A, the
anion derived from
5R-(tert-butyl-dimethyl-silanyloxymethyl)-pyrrolidin-2-one (20 g,
87 mmol) and NaHMDS (1M in THF, 96 mL, 96 mmol) was alkylated with
7-bromoheptanenitrile (13 mL, 87 mmol). The reaction mixture was
stirred at 60.degree. C. for 24 h and was worked-up analogous to
that described for Preparation 1, Step A. The crude residue was
dissolved in MeOH (350 mL) and 1N HCl (154 mL) was added. The
solution was stirred for 2 h and the volatiles were removed in
vacuo. The aqueous solution was washed with CH.sub.2Cl.sub.2
(3.times.200 mL) and the organic solutions were combined and washed
with brine (1.times.150 mL). The organic solution was dried
(Na.sub.2SO.sub.4), filtered and concentrated. Purification by
medium pressure chromatography (1% MeOH in EtOAc to 4% MeOH in
EtOAc) provided
7-(2R-hydroxymethyl-5-oxo-pyrrolidin-1-yl)-heptanenitrile (10.3 g).
.sup.1H NMR (CDCl.sub.3) .delta. 3.76 (dd, 1H), 3.62 (m, 3H), 2.97
(m, 1H), 2.43 (m, 1H), 2.33-1.94 (m, 5H), 1.92 (m, 1H), 1.66-1.41
(m, 6H), 1.30 (m, 2H); MS 225.3 (M+1).
Preparation 4
4-(3-Bromo-propyl)-benzoic acid methyl ester
Step A: 4-(3-Hydroxy-prop-1-ynyl)-benzoic acid methyl ester. To a
solution of methyl 4-iodobenzoate (20 g, 76 mmol), propargyl
alcohol (5.55 g, 99.0 mmol) and triethylamine (20 mL) in
acetonitrile (200 mL) was added
dichlorobis(triphenylphosphine)palladium(II) (1.55 g, 2.21 mmol),
followed by CuI (454 mg, 2.38 mmol). The reaction mixture was
stirred at room temperature for 24 h. Water was added and the
aqueous solution was washed with EtOAc (3.times.). The organic
solutions were combined, dried (MgSO.sub.4), filtered and
concentrated. Purification by medium pressure chromatography (9:1
hexanes:EtOAc to 4:1 hexanes:EtOAc) provided
4-(3-hydroxy-prop-1-ynyl)-benzoic acid methyl ester (12.65 g).
Step B: 4-(3-Hydroxy-propyl)-benzoic acid methyl ester. A solution
of 4-(3-hydroxy-prop-1-ynyl)-benzoic acid methyl ester (12.65 g) in
EtOAc (75 mL) and MeOH (75 mL) was hydrogenated at 50 psi on a Parr
shaker in the presence of 10% palladium on carbon (2 g) for 24 h.
The catalyst was removed by filtration through Celite.RTM. and the
filtrate was concentrated. The reaction was repeated by adding 10%
palladium on carbon (2 g) and hydrogenating on a Parr shaker for 24
h. After filtering through Celite.RTM., the solution was
concentrated in vacuo to provide 4-(3-hydroxy-propyl)-benzoic acid
methyl ester (11.98 g).
Step C: 4-(3-Bromo-propyl)-benzoic acid methyl ester. A solution of
4-(3-hydroxy-propyl)-benzoic acid methyl ester (11.98 g) and
1,1'-carbonyldiimidazole (9.0 g, 55.50 mmol) in CH.sub.3CN (200 mL)
was stirred at room temperature for 1.5 h. Allyl bromide (20 mL)
was added and the reaction mixture was heated under reflux for 20
h. The reaction mixture was cooled to room temperature and
saturated aqueous NaHCO.sub.3 was added. The aqueous solution was
washed with EtOAc (3.times.) and the organic solutions were
combined, dried (MgSO.sub.4), filtered and concentrated.
Purification by medium pressure chromatography (9:1 hexanes:EtOAc)
provided the title compound of Preparation 4.
Preparation 5
2-[3-(2R-Hydroxymethyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiazole-4-carboxyli-
c acid ethyl ester
Step A: 2-Bromo-thiazole-4-carboxylic acid ethyl ester. A cold
solution of sodium nitrite (228 mg, 3.31 mmol) in water (2.0 mL)
was added dropwise to a mixture of 2-amino-thiazole-4-carboxylic
acid ethyl ester (J. Am. Chem. Soc., 1946, 68, 266) (500 mg, 2.90
mmol), CuSO.sub.4 pentahydrate (2.100 g, 8.41 mmol), NaBr (1.134 g,
11.02 mmol), H.sub.2SO.sub.4 (3.0 mL) and water (3.0 mL) at
-5.degree. C. to 0.degree. C. The reaction mixture was stirred at
0.degree. C. for 20 minutes and at room temperature for 1 h. The
reaction mixture was adjusted to pH 9 with 1N NaOH (105 mL) and the
aqueous solution was washed with CHCl.sub.3 (4.times.50 mL). The
organic solutions were combined, dried (MgSO.sub.4), filtered and
concentrated. Purification by medium pressure chromatography (39:1
hexanes:EtOAc to 19:1 hexanes:EtOAc) provided
2-bromo-thiazole-4-carboxylic acid ethyl ester (257 mg).
Step B:
2-{3-[2R-(tert-Butyl-dimethyl-silanyloxymethyl)-5-oxo-pyrrolidin--
1-yl]-prop-1-ynyl}-thiazole-4-carboxylic acid ethyl ester.
Substituting the appropriate starting materials, the compound of
Step B was prepared using an analogous procedure to that described
for Preparation 4, Step A using tetrakis(triphenylphosphine)
palladium(0) and copper (I) iodide, CuI as catalysts.
Step C:
2-{3-[2R-(tert-Butyl-dimethyl-silanyloxymethyl)-5-oxo-pyrrolidin--
1-yl]-propyl}-thiazole-4-carboxylic acid ethyl ester. Substituting
the appropriate starting materials, the compound of Step C was
prepared using an analogous procedure to that described for
Preparation 4, Step B.
Step D:
2-[3-(2R-Hydroxymethyl-5-oxo-pyrrolidin-1-yl)-propyl]-thiazole-4--
carboxylic acid ethyl ester. To a solution of
2-{3-[2R-(tert-butyl-dimethyl-silanyloxymethyl)-5-oxo-pyrrolidin-1-yl]-pr-
opyl}-thiazole-4-carboxylic acid ethyl ester (306 mg, 0.717 mmol)
in THF (20 mL) at 0.degree. C. was slowly added Bu.sub.4NF (1M in
THF, 1.1 mL, 1.1 mmol). The reaction mixture was warmed to room
temperature and was stirred for 2 h. Aqueous saturated NaHCO.sub.3
was added and the volatiles were concentrated in vacuo. The aqueous
solution was washed with CHCl.sub.3 (4.times.10 mL). The organic
solutions were combined, dried (MgSO.sub.4), filtered and
concentrated to provide the title compound of Preparation 5 (225
mg).
Preparation 6
[3-(4-Fluoro-3-methyl-phenyl)-2-oxo-propyl]-phosphonic acid diethyl
ester
Step A: [3-(4-Fluoro-3-methyl-phenyl)-2-hydroxy-propyl]-phosphonic
acid diethyl ester. To a solution of
4-fluoro-3-methylphenylmagnesium bromide (0.5M in Et.sub.2O, 15.5
mL, 7.75 mmol) in THF (10 mL) at -30.degree. C. was added copper
(I) iodide, CuI (196 mg, 1.03 mmol) and the reaction mixture was
stirred for 10 minutes. The reaction mixture was warmed to
-15.degree. C. and oxiranylmethyl-phosphonic acid diethyl ester (1
g, 5.2 mmol) in THF (10 mL) was added. The reaction mixture was
stirred at 0.degree. C. for 2 h. Saturated aqueous ammonium
chloride was added and the product was extracted into EtOAc. The
organic solution was dried (MgSO.sub.4), filtered and concentrated.
Purification by medium pressure chromatography (20% EtOAc in
hexanes to 70% EtOAc in hexanes) provided
[3-(4-fluoro-3-methyl-phenyl)-2-hydroxy-propyl]-phosphonic acid
diethyl ester (1.37 g).
Step B: [3-(4-Fluoro-3-methyl-phenyl)-2-oxo-propyl]-phosphonic acid
diethyl ester. To a solution of
[3-(4-fluoro-3-methyl-phenyl)-2-hydroxy-propyl]-phosphonic acid
diethyl ester (1.37 g, 4.51 mmol) in CH.sub.2Cl.sub.2 (30 mL) was
added Dess-Martin reagent (Dess-Martin periodinane, Aldrich
Chemical Co., Milwaukee, Wis., 2.10 g, 4.96 mmol). The reaction
mixture was stirred at room temperature for 2 h and additional
CH.sub.2Cl.sub.2 was added. The organic solution was washed with
NaHCO.sub.3 (2 times) and once with brine. The organic solution was
dried (MgSO.sub.4), filtered and concentrated. Purification by
medium pressure chromatography (20% EtOAc in hexanes to 70% EtOAc
in hexanes) provided the title compound of Preparation 6 (1.1
g).
Preparation 7
[3-(3-Methoxymethyl-phenyl)-2-oxo-propyl]-phosphonic acid diethyl
ester
Substituting the appropriate starting materials, the title compound
of Preparation 7 was prepared following an analogous procedure to
that described for Preparation 6.
Preparation 8
[3-(4-Ethyl-phenyl)-2-oxo-propyl]-phosphonic acid diethyl ester
Substituting the appropriate starting materials, the title compound
of Preparation 8 was prepared following an analogous procedure to
that described for Preparation 6.
Preparation 9
{3-[3-(2-Methoxy-ethyl)-phenyl]-2-oxo-propyl}-phosphonic acid
diethyl ester
Substituting the appropriate starting materials, the title compound
of Preparation 9 was prepared following an analogous procedure to
that described for Preparation 6.
Preparation 10
[2-Oxo-3-(3-trifluoromethyl-phenyl)-propyl]-phosphonic acid
dimethyl ester
Step A: N-Methoxy-N-methyl-2-(3-trifluoromethyl-phenyl)-acetamide.
To a solution of N,O-dimethylhydroxylamine hydrochloride (1.577 g,
16.2 mmol) in DMF (25 mL) and CH.sub.2Cl.sub.2 (25 mL) at 0.degree.
C. was added triethylamine (2.25 mL). After stirring for 5 minutes,
3-trifluoromethylphenyl acetic acid (3.0 g, 14.7 mmol), HOBT (3.177
g, 23.5 mmol), and EDC (3.10 g, 16.2 mmol) were added. The reaction
mixture was stirred at room temperature for 18 h and was
concentrated in vacuo. The residue was diluted with EtOAc and the
organic solution was washed consecutively with 1N NaOH (2 times),
water, and brine. The organic solution was dried (MgSO.sub.4),
filtered and concentrated in vacuo. Medium pressure chromatography
(20% EtOAc in hexanes to 50% EtOAc in hexanes) provided
N-methoxy-N-methyl-2-(3-trifluoromethyl-phenyl)-acetamide.
Step B: [2-Oxo-3-(3-trifluoromethyl-phenyl)-propyl]-phosphonic acid
dimethyl ester. To a solution of dimethyl methylphosphonate (9.4 g,
75.8 mmol) in toluene (80 mL) at -78.degree. C. was slowly added
n-BuLi (2.5M in hexanes, 28 mL, 70 mmol). The reaction mixture was
stirred for 1 h and a solution of
N-methoxy-N-methyl-2-(3-trifluoromethyl-phenyl)-acetamide (14.39 g)
in toluene (50 mL) was slowly added. The reaction mixture was
stirred for 2.5 h and AcOH (40 mL) was added. The reaction mixture
was warmed to room temperature and water was added. The organic
layer was washed with water followed by brine. The organic solution
was dried (MgSO.sub.4), filtered and concentrated in vacuo. Medium
pressure chromatography (CH.sub.2Cl.sub.2 to 2% MeOH in
CH.sub.2Cl.sub.2) provided the title compound of Preparation 10
(9.37 g). .sup.1H NMR (CDCl.sub.3) .delta.7.52 (m, 1H), 7.44 (m,
2H), 7.37 (m, 1H), 3.96 (s, 2H), 3.87 (s, 3H), 3.76 (s, 3H), 3.12
(d, 2H).
Preparation 11
[3-(3-Chloro-phenyl)-2-oxo-propyl]-phosphonic acid dimethyl
ester
Substituting the appropriate starting materials, the title compound
of Preparation 11 was prepared following an analogous procedure to
that described for Preparation 10.
Preparation 12
[3-(3-Bromo-phenyl)-2-oxo-propyl]-phosphonic acid dimethyl
ester
Substituting the appropriate starting materials, the title compound
of Preparation 12 was prepared following an analogous procedure to
that described for Preparation 10.
Preparation 13
[2-Oxo-3-(3-trifluoromethoxy-phenyl)-propyl]-phosphonic acid
dimethyl ester
Substituting the appropriate starting materials, the title compound
of Preparation 13 was prepared following an analogous procedure to
that described for Preparation 10. MS 327.1 (M+1), 325.1 (M-1).
Preparation 14
[3-(3-Chloro-phenyl)-2-oxo-propyl]-phosphonic acid dimethyl
ester
To a solution of dimethyl methylphosphonate (17.93 g, 144 mmol) in
THF (270 mL) at -78.degree. C. was slowly added n-BuLi (2.5M, 64.2
mL, 160.6 mmol). The reaction mixture was stirred for 1 h and
(3-chloro-phenyl)-acetic acid methyl ester (26.93 g, 146 mmol) was
slowly added. The reaction mixture was allowed to warm to room
temperature and was stirred for 24 h. Acetic acid (15 mL) was added
and the volatiles were removed in vacuo. The residue was diluted
with CH.sub.2Cl.sub.2 and the organic solution was washed carefully
with saturated aqueous NaHCO.sub.3 (3 times). The organic layer was
dried (MgSO.sub.4), filtered and concentrated in vacuo.
Purification by medium pressure chromatography (20% EtOAc in
hexanes to EtOAc) provided the title compound (9.28 g).
Preparations 15-24
Substituting the appropriate starting materials, the following
phosphonates (Preparations 15-24) were prepared in an analogous
fashion to the procedure described for Preparation 14.
Preparation 15: [3-(3-Fluoro-phenyl)-2-oxo-propyl]-phosphonic acid
dimethyl ester
Preparation 16: [3-(4-Fluoro-phenyl)-2-oxo-propyl]-phosphonic acid
dimethyl ester
Preparation 17: [3-(4-Chloro-phenyl)-2-oxo-propyl]-phosphonic acid
dimethyl ester
Preparation 18: (3-Naphthalen-2-yl-2-oxo-propyl)-phosphonic acid
dimethyl ester
Preparation 19: (2-Oxo-3-thiophen-2-yl-propyl)-phosphonic acid
dimethyl ester
Preparation 20: (3-Cyclohexyl-2-oxo-propyl)-phosphonic acid
dimethyl ester
Preparation 21: (2-Oxo-3-phenyl-propyl)-phosphonic acid dimethyl
ester
Preparation 22: (3-Benzo[1,3]dioxol-5-yl-2-oxo-propyl)-phosphonic
acid dimethyl ester
Preparation 23: [2-Oxo-3-(3-phenoxy-phenyl)-propyl]-phosphonic acid
dimethyl ester
Preparation 24:
[2-Oxo-3-(2-trifluoromethyl-phenyl)-propyl]-phosphonic acid
dimethyl ester
Preparation 25
(3-Biphenyl-3-yl-2-oxo-propyl)-phosphonic acid dimethyl ester
Step A: Biphenyl-3-yl-acetic acid methyl ester. A mixture of
phenylboronic acid (1.000 g, 8.20 mmol), methyl
3-bromophenylacetate (1.691 g, 7.38 mmol), Na.sub.2CO.sub.3 (1.738
g, 16.4 mmol), tetrakis(triphenylphosphine)palladium(0) (0.474 g,
0.41 mmol), toluene (30 mL), and water (5 mL) was heated under
reflux for 20 h. The reaction mixture was diluted with water (20
mL) and the volatiles were removed in vacuo. The aqueous solution
was washed with EtOAc (4.times.20 mL). The organic solutions were
combined, washed with 1N NaOH (15 mL) followed by water (15 mL).
The organic solution was dried (MgSO.sub.4), filtered and
concentrated in vacuo. Purification by medium pressure
chromatography (79:1 hexanes:EtOAc to 39:1 hexanes:EtOAc) provided
biphenyl-3-yl-acetic acid methyl ester (1.316 g).
Step B: (3-Biphenyl-3-yl-2-oxo-propyl)-phosphonic acid dimethyl
ester. The title compound of Preparation 25 was prepared from
biphenyl-3-yl-acetic acid methyl ester of Step A following an
analogous procedure as described for Preparation 14.
Preparation 26
Tetrahydro-pyrrolizine-3,5-dione
The title compound of Preparation 26 was prepared following the
procedure described in U.S. Pat. No. 4,663,464.
In Vitro Assays
The compounds of Formula I, which are useful in the methods of the
present invention, bind to the prostaglandin E.sub.2 type 4
receptor (EP.sub.4 receptor). The full length coding sequence for
the human EP.sub.1 receptor is made in accordance with the
procedure in Funk et al., Journal of Biological Chemistry, 1993,
268, 26767-26772. The full length rat EP.sub.2 receptor is made in
accordance with the procedure in Nemoto et al., Prostaglandins and
other Lipid Mediators, 1997, 54, 713-725. The full length coding
sequence for the human EP.sub.3 receptor is made in accordance with
the procedure in Regan et al., British Journal of Pharmacology,
1994, 112, 377-385. The full length coding sequence for the rat
EP.sub.4 receptor is made in accordance with the procedure in Sando
et al., Biochem. Biophys. Res. Comm. 1994, 200, 1329-1333. These
full length receptors are used to prepare 293S cells expressing the
human EP.sub.1, rat EP.sub.2, human EP.sub.3 or rat EP.sub.4
receptors.
Human EP.sub.1, Rat EP.sub.2, Human EP.sub.3, Rat EP.sub.4 Receptor
Binding Assay
The full length receptors described above are used to prepare 293S
cells expressing the EP.sub.1, EP.sub.2, EP.sub.3, and EP.sub.4
receptors.
293S cells expressing either the human EP.sub.1, rat EP.sub.2,
human EP.sub.3 or rat EP.sub.4 prostaglandin E.sub.2 receptors are
generated according to methods known to those skilled in the art.
Typically, PCR (polymerase chain reaction) primers corresponding to
the 5' and 3' ends of the published full length receptor are made
according to the well known methods disclosed above and are used in
an RT-PCR (reverse transcriptase-polymerase chain reaction)
reaction using the total RNA from human kidney (for EP.sub.1), rat
kidney (for EP.sub.2), human lung (for EP.sub.3), or rat kidney
(EP.sub.4) as a source. PCR products are cloned by the TA overhang
method into pCR2.1 (Invitrogen Corporation, Carlsbad, Calif.) and
identity of the cloned receptor is confirmed by DNA sequencing. For
expression of the rat EP.sub.2 receptor, the confirmed cDNA is
subcloned into the mammalian expression vector PURpCI, a vector
generated by subcloning the selectable marker for puromycin
resistance into the mammalian expression vector pCI (Promega,
Madison, Wis.)
293S cells are transfected with either the cloned human EP.sub.1 or
EP.sub.3 receptor in pcDNA3 by electroporation. Stable cell lines
expressing either the human EP.sub.1 or EP.sub.3 receptor are
established following selection of transfected cells with G418.
293S cells are transfected with the cloned rat EP.sub.2 receptor in
PURpCi by lipid mediated transfection. Stable cell lines expressing
the rat EP.sub.2 receptor are established following selection of
transfected cells with puromycin. 293S cells are transfected with
the cloned rat EP.sub.4 receptor in pcDNA3 by lipid mediated
transfection. Stable cell lines expressing the rat EP.sub.4
receptor are established following selection of transfected cells
with Geneticin.RTM. (Invitrogen, Carlsbad, Calif.).
Clonal cell lines expressing the maximal number of receptors are
chosen following a whole cell .sup.3H-PGE.sub.2 binding assay using
unlabeled PGE.sub.2 as a competitor.
Membrane Preparation: All operations are performed at 4.degree. C.
Transfected cells expressing either prostaglandin E.sub.2 type 1,
type 2, type 3, or type 4 (EP.sub.1, EP.sub.2, EP.sub.3, or
EP.sub.4, respectively) receptors are harvested and suspended to 2
million cells per ml in Buffer A [50 mM Tris-HCl (pH 7.4), 10 mM
MgCl.sub.2, 1 mM EDTA, 1 mM Pefabloc peptide, (Boehringer Mannheim
Corp., Indianapolis, Ind.), 10 uM Phosporamidon peptide, (Sigma,
St. Louis, Mo.), 1 uM pepstatin A peptide, (Sigma, St. Louis, Mo.),
10 uM elastatinal peptide, (Sigma, St. Louis, Mo.), 100 uM antipain
peptide, (Sigma, St. Louis, Mo.)]. The cells are lysed by
sonification with a Branson Sonifier (Branson Ultrasonics
Corporation, Danbury, Conn.) in 2 fifteen second bursts. Unlysed
cells and debris are removed by centrifugation at 100.times.g for
10 min. Membranes are then harvested by centrifugation at
45,000.times.g for 30 minutes. Pelleted membranes are resuspended
to 3-10 mg protein per ml, protein concentration being determined
of the method of Bradford [Bradford, M., Anal. Biochem. 1976, 72,
248]. Resuspended membranes are then stored frozen at -80.degree.
C. until use.
Binding Assay: Frozen membranes prepared as above are thawed and
diluted to 1 mg protein per ml in Buffer A above. 100 .mu.l of the
cell membrane preparation is mixed with 5 .mu.l of a solution of
test compound of Formula I (diluted in DMSO to a concentration 40
times the desired final concentration) and 95 .mu.l of 3 nM
.sup.3H-prostaglandin E.sub.2 (Amersham, Arlington Heights, Ill.)
in Buffer A. The mixture (200 .mu.L total volume) is incubated for
1 hour at 25.degree. C. The membranes are then recovered by
filtration through type GF/C glass fiber filters (Wallac,
Gaithersburg, Md.) using a Tomtec harvester (Tomtec, Orange,
Conn.). The membranes with bound .sup.3H-prostaglandin E.sub.2 are
trapped by the filter, while the buffer and unbound
.sup.3H-prostaglandin E.sub.2 pass through the filter into waste.
Each sample is then washed 3 times with 3 ml of [50 mM Tris-HCl (pH
7.4), 10 mM MgCl.sub.2, 1 mM EDTA]. The filters are then dried by
heating in a microwave oven. To determine the amount of
.sup.3H-prostaglandin bound to the membranes, the dried filters are
placed into plastic bags with scintillation fluid and counted in a
LKB 1205 Betaplate reader (Wallac, Gaithersburg, Md.). IC.sub.50s
are determined from the concentration of test compound required to
displace 50% of the specifically bound .sup.3H-prostaglandin
E.sub.2.
Determination of Cyclic AMP Elevation in 293S Cell Lines Stably
Overexpressing Recombinant Rat EP.sub.4 Receptors Assay
cDNA representing the complete open reading frame of the rat
EP.sub.4 receptor is generated by reverse transcriptase polymerase
chain reaction using oligonucleotide primers based on published
sequences. The full length coding sequence for the rat EP.sub.4
receptor is made in accordance with the procedure in Sando et al.,
Biochem. Biophys. Res. Comm. 1994, 200, 1329-1333, and RNA from rat
kidney (EP.sub.4) as templates. 293S cells are transfected with the
cloned rat EP.sub.4 receptor in pcDNA3 by lipid mediated
transfection. Stable cell lines expressing the rat EP.sub.4
receptor are established following selection of transfected cells
with Geneticin.RTM. (Invitrogen Corporation, Carlsbad, Calif.).
Clonal cell lines expressing the maximal number of receptors are
chosen following a whole cell .sup.3H-PGE.sub.2 binding assay using
unlabeled PGE.sub.2 as a competitor. Transfectants demonstrating
high levels of specific [.sup.3H]PGE.sub.2 binding are further
characterized by Scatchard analysis to determine B.sub.max and
K.sub.ds for PGE.sub.2. The lines selected for compound screening
have approximately 256,400 receptors per cell and a K.sub.d=2.9 nm
for PGE.sub.2 (EP.sub.4). Constitutive expression of the receptor
in parental 293-S cells is negligible. A stable cell line
containing the rat EP.sub.4 receptor is grown in Dulbecco's
Mosified Eagle Medium/F12 (DMEM/F12) containing 10% fetal bovine
serum and G418 (500 .mu.g/ml) to 80% confluency.
cAMP responses in the 293-S/EP.sub.4 lines are determined by
detaching cells from culture flasks in 1 ml of calcium (Ca++) and
magnesium (Mg++) deficient phosphate buffered saline (PBS) via
vigorous pounding and then rinsing the cells with calcium (Ca++)
and magnesium (Mg++) deficient phosphate buffered saline (PBS). The
cells are resuspended in MEM (Minimum Essential Medium), 1% BSA
(bovine serum albumin), 50 mM HEPES
(N-[2-Hydroxyethyl]piperazine-N'-[2-ethanesulfonic acid]) at
37.degree. C. The cell suspension is counted on a hemacytometer and
diluted by adding MEM (Minimum Essential Medium) to a final
concentration of 1.times.10.sup.6 cells/ml, and adding
3-isobutyl-1-methylxanthine (IBMX) to a final concentration of 1
mM. 200 microliters of cell suspension is immediately aliquoted
into individual tubes and incubated for 10 minutes, uncovered, at
37.degree. C., 5% CO.sub.2, 95% relative humidity. The compound of
Formula I to be tested in either dimethylsulfoxide (DMSO) or
ethanol is then added to cells at 1:100 dilutions such that the
final DMSO or ethanol concentration is 1%. Typically, the cells are
treated with 6-8 different concentrations (in 1 log increments,
such as those described below) of the compound of Formula I.
Typical concentrations of the compound of Formula I in this assay
are between 10.sup.-5M to 10.sup.-10M. For example, a six point
compound dose response assay tests the compound of Formula I at
concentrations of 10.sup.-5M, 10.sup.-6M, 10.sup.-7M, 10.sup.-8M,
10.sup.-9M and 10.sup.-10M. Immediately after adding the test
compound, the tubes are covered, mixed by inverting two times, and
incubated at 37.degree. C. for 12 minutes. Samples are then lysed
by incubation at 100.degree. C. for 10 minutes and immediately
cooled on ice for 5 minutes to approximately 4.degree. C. Cellular
debris is pelleted by centrifugation at 3500.times.g for 5 minutes
at approximately 4.degree. C., and cleared lysates are transferred
to fresh tubes. cAMP concentrations are determined using a
commercially available .sup.125I-cAMP radioimmunoassay (RIA) kit
(NEK-033, Perkin-Elmer Life Sciences, Inc., Boston, Mass.). The
cleared lysates are diluted 1:100 in cAMP RIA assay buffer
(included in kit) and centrifuged again. 50 microliters of the
resulting supernatant is transferred to a 12.times.75 mm glass tube
and data is collected by scintillation counting using a Wallac
Cobra II Gamma Counter (Perkin-Elmer Wallac, Inc., Gaithersburg,
Md.). EC.sub.50 calculations are performed on a calculator using
linear regression analysis on the linear portion of the dose
response curves or using Data Fitter.
In Vivo Assays
The selective EP.sub.4 receptor agonists of Formula I can be
evaluated in various in vivo liver failure models known in the art,
such as the in vivo rat liver failure model as disclosed by Kasai,
K. et al. in Gastroenterology 2001, 120 (suppl. 1), A-541.
In Vivo Acute Liver Injury Model
Methods: Acute liver failure in rats can be induced by
intraperitoneal injection of one of carbon tetrachloride
(CCl.sub.4, 1 mg/kg), dimethylnitrosamine (DMN, 50 mg/kg),
D-galactosamine (D-gal, 1 g/kg), or D-galactosamine with
lipopolysaccharide (LPS), (D-gal, 1 g/kg; LPS 100 .mu.g/kg).
Immediately following the intaperitoneal injection of carbon
tetrachloride, dimethyinitrosamine, D-galactosamine, or
D-galactosamine with lipopolysaccharide, the test compound of
Formula I or saline (as control) is administered. The test compound
(a selective EP.sub.4 receptor agonist of Formula I) can be
administered at various doses such as 0.01, 0.05, 0.1 or 0.2 mg/kg.
24 hours after administration of the test compound of Formula I,
the liver can be removed for histology and serum can be obtained
for determination of total bilirubin (T-bil), aspartate
aminotransferase (AST), and alanine aminotransferase (ALT). Massive
hepatic necrosis with marked elevations in the levels of T-bil,
AST, and ALT was observed in the saline treated control group. The
effectiveness of the test compound in the above models can be
determined by comparison of histology and serum results obtained
for the animals treated with the test compound with the
corresponding results from the saline control group.
The following in vivo anesthetized rabbit model is used in order to
demonstrate the hypotensive effect of the compounds of Formula I
(e.g., Example 3).
In Vivo Rabbit Model
Methods: New Zealand White male rabbits (3-4 kg) are anesthetized
with sodium pentobarbital (30 mg/kg, i.v.) and a surgical plane of
anesthesia is maintained by a continuous infusion of sodium
pentobarbital (16 mg/kg/hr) via an ear vein catheter. A tracheotomy
is performed through a ventral midline cervical incision and the
rabbits are ventilated with 100% oxygen using a positive pressure
ventilator. Body temperature is maintained at 38.5.degree. C. using
a heating pad connected to a YSI temperature controller model 72
(Yellow Springs Instruments, Yellow Springs, Md.). Fluid-filled
catheters are placed in the right jugular vein (for intravenous
drug administration) and in the right carotid artery for arterial
pressure monitoring and for blood gas analysis using a model 248
blood gas analyzer (Bayer Diagnostics, Norwood, Mass.). The
ventilator is adjusted as needed to maintain blood pH and pCO.sub.2
within normal physiological ranges for rabbits. Arterial pressure
is measured using a strain gauge transducer (Spectromed, Oxnard,
Calif.), previously calibrated using a mercury manometer,
positioned at the level of the heart and connected to the arterial
catheter. Arterial pressure signals are digitized at 500 Hz and
analyzed using a Po--Ne-Mah Data Acquisition System (Gould
Instrument Systems, Valley View, Ohio) to obtain mean arterial
pressure and heart rate values. Baseline values are collected when
mean arterial pressure and heart rate have stabilized. The test
compound (Compound of Formula I) is then administered either as a
subcutaneous (SC) bolus or as an intravenous (IV) infusion. For
subcutaneous (SC) dosing the test compound can be dissolved in an
appropriate vehicle such as 5% ethanol in water (5% EtOH 95%
H.sub.2O), while for intravenous dosing the test compound can be
dissolved in an appropriate vehicle such as 0.9% normal saline.
Arterial pressure and heart rate are monitored continuously for 4
hours following dosing of the test compound or for the duration of
a continuous 4 hour infusion of the test compound. Blood is sampled
after dosing or during the infusion of the test compound to
determine plasma concentrations of the test compounds.
Data Analysis: Data are presented as mean values. The hemodynamic
data (heart rate and mean arterial pressure) are collected over 4
hours post-dosing in all groups and the reported value is the
average value over the 5-minute interval prior to the selected
time.
The following in vivo primate model is used in order to demonstrate
the hypotensive effect of the compounds of Formula I in primates
(e.g., Example 4).
In Vivo Primate Model
Methods: Adult M. fascicularis primates (6-8 kg) that have been
previously instrumented with subcutaneous vascular access ports in
the descending thoracic aorta and conditioned to sit quietly in
specially designed primate-restraining chairs are used. All
primates are fasted for 12-18 hours prior to the experiment. On the
day of the experiment, with the primates restrained in the chairs,
a strain gauge pressure transducer (Spectromed, Oxnard, Calif.),
previously calibrated using a mercury manometer, is positioned at
the level of the heart and connected to the vascular access port to
measure arterial pressure. The primates are allowed to acclimate to
the chair for at least one hour. Arterial pressure signals are
digitized at 500 Hz and continuously recorded throughout the
experiment and analyzed using a Po--Ne-Mah Data Acquisition System
(Gould Instrument Systems, Valley View, Ohio) to obtain the
measurements of mean arterial pressure and heart rate. Baseline
values are collected when the primates are sitting calmly and when
mean arterial pressure and heart rate have stabilized. The test
compound (Compound of Formula I) is then administered as a
subcutaneous (SC) bolus of a solution of the test compound in an
appropriate vehicle such as 5% ethanol in water (5% EtOH 95%
H.sub.2O). The solution of test compound or vehicle is filtered
through a 0.22 micron filter prior to injection and a typical
dosing volume is 0.2 ml/kg. Arterial pressure and heart rate are
monitored continuously for 4 hours following dosing of the test
compound and are recorded at selected time intervals for data
comparison (vehicle vs test compound). Blood samples (1.5 ml) are
withdrawn to determine plasma concentrations of the test compound
and withdrawn blood is immediately replaced with 0.9% sterile
saline to maintain blood volume.
Data Analysis: Data are presented as mean values. The hemodynamic
data (heart rate and mean arterial pressure) are collected over 4
hours post-dosing in all groups and the reported value is the
average value over the 5-minute interval prior to the selected
time.
EXAMPLE 1
The Human EP.sub.1, Rat EP.sub.2, Human EP.sub.3, Rat EP.sub.4
Receptor Binding Assay, described hereinabove, was used in order to
demonstrate the binding of
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid (Compound 3M) to the
human EP.sub.1, rat EP.sub.2, human EP.sub.3, and rat EP.sub.4
receptors.
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid (Compound 3M) was run in
the assay described and the following IC.sub.50s were obtained.
IC.sub.50s: human EP.sub.1 receptor, >1000 nm; rat EP.sub.2
receptor, 463 nm; human EP.sub.3 receptor, >1000 nm; and rat
EP.sub.4 receptor, 11 nm. These results show that
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid (Compound 3M) binds
selectively to the rat EP.sub.4 receptor in the assay
described.
EXAMPLE 2
The cyclic AMP Elevation in 293S Cell Lines Stably Overexpressing
Recombinant Rat EP.sub.4 Receptors Assay, described hereinabove,
was used in order to demonstrate the effect of
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid (Compound 3M).
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid (Compound 3M) was run in
the assay described and an EC.sub.50 of 0.6 nm was obtained.
EXAMPLE 3
The in vivo rabbit model, described hereinabove, was used in order
to demonstrate the hypotensive effect of
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid (Compound 3M, sodium
salt), at the dosages described below.
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid sodium salt (Compound
3M, sodium salt) was administered according to the previously
described method either as a subcutaneous (SC) bolus (in 5% ethanol
in water) or as an intravenous (IV) infusion (in 0.9% normal
saline).
Compound: The test compound,
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid sodium salt (Compound
3M, sodium salt), was adjusted for active compound (mgA) and
dissolved in the stated vehicle at the following concentrations:
For Group A, 5 mgA/ml in 5% EtOH: 95% H.sub.2O; for Group B;
approximately 0.1 mgA/ml in 0.9% normal saline; for Group C;
approximately 0.01 mgA/ml in 0.9% normal saline (tenfold dilution
of the Group B solution with 0.9% saline); and for Group D;
approximately 0.001 mgA/ml in 0.9% normal saline (tenfold dilution
of the Group C solution with 0.9% saline). Thus, dosing volumes
were 0.2 ml/kg (Group A) subcutaneously or 5 ml/h (Groups B, C, and
D) as an intravenous (IV) infusion. The term "mgA" means the number
of milligrams adjusted for active compound (e.g. corrected for salt
etc.).
Dosing: Four groups (Groups A, B, C, and D) of two rabbits each
were dosed as follows.
Group A (2 rabbits) received Compound 3M, as its sodium salt, in
vehicle (5% EtOH:95% H.sub.2O) as a subcutaneous (SC) bolus, at 1
mgA/kg (0.2 ml/kg of the 5 mgA/ml solution described above).
Group B (2 rabbits) received Compound 3M, as its sodium salt, in
vehicle (approximately 0.1 mgA/ml in 0.9% normal saline solution,
described above), infused intravenously (IV) at 167 .mu.g/kg/hr for
4 hours at 5 ml/hr.
Group C (2 rabbits) received Compound 3M, as its sodium salt, in
vehicle (approximately 0.01 mgA/ml in 0.9% normal saline solution,
described above) infused IV at 16.7 .mu.g/kg/hr for 4 hours at 5
ml/hr.
Group D (2 rabbits) received Compound 3M, as its sodium salt, in
vehicle (approximately 0.001 mgA/ml in 0.9% normal saline solution,
described above) infused IV at 1.67 .mu.g/kg/hr for 4 hours at 5
ml/hr.
Data analysis was carried out as described in the general in vivo
rabbit model procedure, hereinabove, and is provided for Groups A,
B, C, and D in Tables 1-4, respectively.
Results:
Group A: The administration of Compound 3M sodium salt at 1 mgA/kg
SC, as described above, caused an increase in heart rate and a
decrease in mean arterial pressure (hypotension) that was rapid in
onset (<2 minutes) and was sustained over the entire 4 hour
post-dose interval (see Table 1).
Group B: The administration of Compound 3M sodium salt at 167
.mu.g/kg/h IV, as described above, caused an increase in heart rate
and a decrease in mean arterial pressure (hypotension) that was
rapid in onset (<2 minutes) and was sustained over the entire 4
hour post-dose interval (see Table 2).
Group C: The administration of Compound 3M sodium salt at 16.7
.mu.g/kg/h IV, as described above, caused a slight increase in the
heart rate and a slight decrease in mean arterial pressure
(hypotension), (see Table 3).
Group D: The administration of Compound 3M sodium salt at 1.67
.mu.g/kg/h, as described above, resulted in no significant change
in either heart rate or mean arterial pressure over the duration of
the 4 hour IV infusion (no significant hemodynamic effects were
observed), (see Table 4).
EXAMPLE 4
The in vivo primate model, described hereinabove, was used in order
to demonstrate the hypotensive effect of
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid sodium salt (Compound
3M, sodium salt), at the dosages described below. The test compound
(Compound 3M, sodium salt) was administered subcutaneously (SC) as
a solution in 5% ethanol in water (5% EtOH:95% H.sub.2O). The dose
volume for compound solution or vehicle control was 0.2 ml/kg
administered as a SC bolus.
Compound: The test compound,
5-(3-{2S-[3R-Hydroxy-4-(3-trifluoromethyl-phenyl)-butyl]-5-oxo-pyrrolidin-
-1-yl}-propyl)-thiophene-2-carboxylic acid sodium salt (Compound
3M, sodium salt), was adjusted for active compound (mgA) and
dissolved in vehicle (5% EtOH, 95% H.sub.2O) at a concentration of
5 mgA/ml for Group A, and 0.5 mgA/ml for Group C. Group B received
vehicle (5% EtOH, 95% H.sub.2O) as a control.
Dosing: Three groups of monkeys (A, B, and C) were dosed as
follows.
Group A: Three male monkeys received Compound 3M, as its sodium
salt, in vehicle (5% EtOH:95% H.sub.2O) SC, at 1 mgA/kg (0.2 ml/kg
of the 5 mgA/ml solution described above).
Group B: Three male monkeys received vehicle (5% EtOH:95% H.sub.2O)
at 0.2 ml/kg.
Group C: Two of the previously vehicle-treated monkeys (from Group
B) received Compound 3M, as its sodium salt, in vehicle (5%
EtOH:95% H.sub.2O) SC, at 0.1 mgA/kg (0.2 ml/kg of the 0.5 mgA/ml
solution described above).
Data analysis was carried out as described in the general in vivo
primate model procedure, hereinabove, and is provided for Groups C
and B in Tables 5-6, respectively.
Results:
Group A: The administration of Compound 3M, sodium salt, at 1
mgA/kg SC, in three monkeys as described above, resulted in a
transient increase in heart rate and a decrease in mean arterial
pressure (hypotension) that was rapid in onset (<2 minutes) and
was sustained over the 4 hours post-dose. The maximum hypotensive
effect could not be determined as treatment, including tilting to
reclining position, was required for all three monkeys to maintain
the mean arterial pressure above 40 mmHg (considered the minimum
required for organ perfusion). The monkeys were gradually returned
to an upright seated position over the course of the study as their
mean arterial pressure allowed (over 30-210 minutes).
Group B: The administration of vehicle, (5% EtOH:95% H.sub.2O) at
0.2 ml/kg, SC, in three monkeys did not substantially affect mean
arterial pressure (MAP) or heart rate (HR) over the 4 hours post
dose (see Table 6).
Group C: The administration of Compound 3M, sodium salt, at 0.1
mgA/kg SC, in 2 monkeys as described above, resulted in a transient
increase in heart rate that returned toward normal but leveled off
and remained elevated over the 4 hours post-dose. The
administration of Compound 3M, sodium salt, at 0.1 mg/kg SC also
caused a decrease in mean arterial pressure (hypotension) that was
rapid in onset (<4 minutes) and was sustained over the 4 hours
post-dose (see Table 5). Initially, the mean arterial pressure
leveled off above 40 mmHg for both monkeys. However, 1 monkey
required a full tilt to a reclining position at 75 minutes
post-dose, when his pressure fell below 40 mmHg, and was returned
to a full upright position by 180 minutes.
TABLES
Tables 1-4 provide data from the in vivo rabbit model and Tables
5-6 provide data from the in vivo primate model, both of which are
described hereinabove. In the tables time is given in minutes, mean
arterial pressure (MAP) is in mm Hg, and heart rate (HR) is in
beats/minute. Baseline MAP and HR values are average values over
the 5-minute interval prior to dosing.
TABLE-US-00001 TABLE 1 Time Baseline 5 10 15 30 60 90 120 150 180
210 240 MAP 92 69 59 60 59 59 59 58 57 60 61 65 HR 243 268 283 285
284 288 284 278 275 273 273 273
TABLE-US-00002 TABLE 2 Time Baseline 5 10 15 30 60 90 120 150 180
210 240 MAP 87 67 67 69 68 67 57 55 52 53 56 55 HR 260 296 296 296
291 293 282 282 279 285 284 297
TABLE-US-00003 TABLE 3 Time Baseline 5 10 15 30 60 90 120 150 180
210 240 MAP 95 91 86 84 84 85 83 85 85 85 89 89 HR 291 289 299 302
306 300 301 304 301 298 307 306
TABLE-US-00004 TABLE 4 Time Baseline 5 10 15 30 60 90 120 150 180
210 240 MAP 89 85 87 85 85 87 87 85 87 87 85 92 HR 260 258 260 262
262 255 253 246 246 246 247 257
TABLE-US-00005 TABLE 5 Time Baseline 5 10 15 30 60 90 120 150 180
210 240 MAP 101 91 57 51 53 58 58 62 65 66 71 79 HR 161 203 206 187
190 180 180 183 187 187 191 193
TABLE-US-00006 TABLE 6 Time Baseline 5 10 15 30 60 90 120 150 180
210 240 MAP 114 115 112 111 112 112 111 110 111 113 115 112 HR 177
179 176 173 179 179 178 181 182 184 191 188
* * * * *